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Registration Dossier
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EC number: 918-668-5 | CAS number: 128601-23-0
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Hydrocarbons, C9 Aromatics:
Genetic Toxicity in vitro - Bacterial reverse mutation assay (OECD TG 471): Negative
Genetic Toxicity in vitro - In vitro Mammalian Chromosome Aberration Test (OECD TG 473): Negative
Genetic Toxicity in vitro - In vitro Mammalian Cell Gene Mutation Test (OECD TG 476): Negative
Genetic Toxicity in vitro - In vitro Sister Chromatid Exchange Assay in Mammalian Cells (OECD TG 479): Negative
Hydrocarbons, C10 Aromatics:
Genetic Toxicity in vitro - Bacterial reverse mutation assay (OECD TG 471): Negative
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:
- 1998/11/05-1999/01/25
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: According to OECD 471 guidelines. GLP.
- 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 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 liver fractions from Aroclor exposed rats
- Test concentrations with justification for top dose:
- Tests (done in triplicate) with and without Metabolic Activation: 0, 6.25, 12.5, 25, 50, 100, 200 ug/plate
Positive controls:
TA 1535 (S9-: sodium azide 1.0 ug/plate) (S9+: 2-aminoanthracene: 2.0 ug/plate)
TA 1537 (S9-: 9-aminoacridine 80 ug/plate) (S9+: benzo(a)pyrene: 4.0 ug/plate)
TA 98 (S9-: 2-nitrofluorene 2.0 ug/plate) (S9+: 2-aminoanthracene: 2.0 ug/plate)
TA 100 (S9-: sodium azide 1.0 ug/plate) (S9+: 2-aminoanthracene: 2.0 ug/plate)
TA102 (S9-: glutaraldehyde 25 ug/plate) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- yes
- Remarks:
- non treated
- Positive controls:
- yes
- Positive control substance:
- other: See Test Concentrations
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar
DURATION
- Exposure duration: 48 hours
NUMBER OF REPLICATIONS:
- triplicate
DETERMINATION OF CYTOTOXICITY
- Method: reduction in the number of revertants and/or clearing of the background lawn of bacterial growth - Evaluation criteria:
- The mutagenicity study is considered valid if the mean colony counts of the control values of the strains are within acceptable ranges, if the positive controls meet the criteria for a positive response and if no more than 5% of the plates are lost through contamination or other unforeseen events.
A test substance is considered to be positive in the bacterial gene mutation test if the mean number of revertant colonies on the test plates increases in a concentration-related manner and/or if a reproducible, two-fold or more increase is observed compared to that on the negative control plates.
A test substance is considered negative in the bacterial gene mutation test if it produces neither a dose-related increase in the mean number of revertant colonies nor a reproducible positive response at any of the test points.
Positive results from the bacterial reverse mutation test indicate that a substance induces point mutations by base substitution for frameshifts in the genome of Salmonella typhimurium. Negative results indicate that under the test conditions, the test substance in not mutagenic. - Statistics:
- The mean plate count and standard deviation for each dose point were determined. Any test value that was equal to or greater than two times the mean value of the concurrent vehicle control was considered to be a positive dose.
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Toxicity was observed at doses above 200 ug/plate with and without S9.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
In all cases, Shellsol AB did not induce any significant changes in the number of revertant colonies. It is concluded in this study that Shellsol AB is not a mutagenic agent. - Executive summary:
Shellsol AB was examined for mutagenic activity in the bacterial reverse mutation test using histidine-requiring Salmonella typhimurium strains TA 1535, 1537, 98, 102,and 100 in the absence and presence of a liver S9 fraction for metabolic activation. Concentrations above 200 ug/plate were found to be cytotoxic and so the test was performed in triplicate using doses of 0, 6.25, 12.5, 25, 50, 100, 200 ug/plate. In all cases, Shellsol AB did not induce any significant changes in the number of revertant colonies. It is concluded in this study that Shellsol AB is not a mutagenic agent.
- 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:
- 06/11/1997-01/13/1997
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP study done according to standard method.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- other: histidine and biotin requiring
- Species / strain / cell type:
- S. typhimurium TA 102
- Additional strain / cell type characteristics:
- other: histidine requiring
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9
- Test concentrations with justification for top dose:
- Range finding: 8, 40, 200, 1000, 5000 µg/plate
Mutation Experiment 1 without metabolic activation: 0.5333, 2.67, 13.3, 66.7, 333.0 µg/plate
Mutation Experiment 1 with metabolic activation: 2.67, 8, 40, 200, 1000 µg/plate
Mutation Experiment 2 without metabolic activation: 31.25, 62.5, 125, 250, 500 µg/plate
Mutation Experiment 2 with metabolic activation, strains TA98 and TA100 initial treatments: 12.5, 25, 50, 100, 200 µg/plate
Mutation Experiment 2 with metabolic activation, strains TA1535, TA1537 and TA102 initial treatments: 62.5, 125, 250, 500, 1000 µg/plate
Mutation Experiment 2 with metabolic activation, strains TA1535 and TA102 repeat treatments: 7.8125, 15.625, 31.25, 62.5, 125, 250 µg/plate
Mutation Experiment 2 with metabolic activation, strain TA1535: 3.90625, 7.8125, 15.625, 31.25, 62.5, 125 µg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- other: not applicable
- Positive controls:
- yes
- Positive control substance:
- other: 2-nitrofluorene, sodium azide, 9-aminoacridine, gluteraldehyde, 2-aminoanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation) - Platings were made by adding 0.1 ml bacterial culture, 0.1 ml test article solution, and 0.5 ml 10% S-9 mix or buffer to 2.5 ml molten agar at 46°C. The plating mixture was rapidly mixed and poured onto Minimal Davis agar plates. After setting, the plates were inverted and incubated for 3 days at 37°C in the dark. Plates were then examined for toxicity. In experiment 2, the test solution was reduced to 0.05 ml due to the toxicity of the solvent, DMSO.
DURATION
- Preincubation period: Experiment 2 - 1 hour
- Exposure duration: 3 days
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):
NUMBER OF REPLICATIONS: 3
OTHER: Colonies were counted using a Seescan Colony Counter or manually when automatic counts could not be obtained. - Evaluation criteria:
- Mean control counts fell within normal ranges.
Positive controls induced clear increases in revertant numbers.
No more than 5% of the plates lost through contamination.
Dunnett's test gave a significant response (p <= 0.01), and the data set showed a significant dose-correlation.
The positive responses were reproducible. - Statistics:
- Individual plate counts were averaged. Dunnett's test was used to calculate significant responses.
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 200, 1000, 5000 µg/plate in TA100
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 200, 1000, 5000 µg/plate in TA100
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 200, 1000, 5000 µg/plate in TA100
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 200, 1000, 5000 µg/plate in TA100
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
It was concluded that the substance did not induce mutation in five strains of Salmonella typhimurium (TA98, TA100, TA1535, TA1537, and TA102 ) when tested under the conditions employed in this study, which included treatment at concentrations up to the lower limit of toxicity, both in the absence and in the presence of rat liver metabolic activation system (S-9). - Executive summary:
A bacterial reverse mutation assay was performed on five strains of Salmonella typhimurium. The study was performed both with (maximum concentration of 1000 microgram/ml) and without metabolic activation (maximum concentration of 500 microgram/ml). After 3 days of exposure to the test substance, the number of mean revertants per plate was calculated. Toxicity was seen at the highest doses. Most notably in the strains TA1535, TA1537, and TA102 during Experiment 2. These strains were retested at lower doses. There was no significant increase in the number of revertants in any of the test strains. There was a significant increase in the number of revertants in the positive control treatment. The study is therefore valid, and 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:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1989
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Acceptable well-documented publication which meets basic scientific principles.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- CHO-K1-BH4 clone
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 from livers of aroclor-induced Sprague-Dawley rats
- Test concentrations with justification for top dose:
- 15.0, 30.1, 45.0, 60.0, 75.0, 90.0 µg/ml without metabolic activation
20.0, 25.0, 37.5, 40.1, 50.0, 60.1, 70.0, 80.2 µg/ml with metabolic activation - Vehicle / solvent:
- DMSO
- Untreated negative controls:
- yes
- Remarks:
- DMSO
- Negative solvent / vehicle controls:
- other: not applicable
- True negative controls:
- other: not applicable
- Positive controls:
- yes
- Positive control substance:
- other: MMC (unactivated cultures) and CP (metabolic activation)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in suspension
DURATION
- Exposure duration: 7 hrs (w/o activation); 2.5 hrs (w/ activation)
- Expression time (cells in growth medium): After 10 hour incubations, metaphase cells were collected by mitotic shake off
- Fixation: methanol:glacial acetic acid (3:1), stained with 5% Giemsa (pH 6.8)
Positive Controls: MMC (unactivated cultures) and CP (metabolic activation) - Evaluation criteria:
- Treatment groups were evaulated for overall chromosomal aberration frequency, the percentage of cells with aberrations, the percentage of cells with more than one aberration, the presence or absence of a dose-response and the complexity of the aberrations.
- Statistics:
- Fisher’s exact test with an adjustment for multiple comparisons to compare the percentage of cells with aberrations in each treatment group with the results from the controls (Sokal and Rohlf, 1981).
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- cytotoxic at 90.2 µg/mL with metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- There were no increases in chromosomal aberrations at any concentration level tested in either the presence or absence of metabolic activation.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
The test material is not clastogenic, however, it is cytotoxic at concentrations of 80.2 µg/ml or above. - Executive summary:
This study assessed the potential of high flash aromatic naphtha to cause chromosome aberrations in Chinese hamster Ovary (CHO) cells. Cells were exposed to concentrations of 15.0, 30.1, 45.0, 60.0, 75.0, 90.0 micrograms/ml without metabolic activation and 20.0, 25.0, 37.5, 40.1, 50.0, 60.1, 70.0, 80.2 micrograms/ml with metabolic activation. Cells were than evaluated for chromosomal aberration frequency, the percentage of cells with aberrations, the percentage of cells with more than one aberration, the presence or absence of a dose-response, and the complexity of the aberrations. There were no increases in chromosomal aberrations at any concentration level tested in either the presence or absence of metabolic activation. The test material is not clastogenic, however, it is cytotoxic at concentrations of 80.2 micrograms/ml or above.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- 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.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Principles of method if other than guideline:
- Method was according to:
Hsie, AW, Brimer, PA, Mitchell, TJ, and Gosslee, DG. (1975) Dose-response relationship for ethyl methanesulfonate-induced mutations at the hypoxyanthine-guanine phosphoribosyl transferase locus in Chinese hamster ovary cells. Somat. Cell Genet. 1:247-261
Hsie, AW, Casciano, DA, Couch, DB, Krahn, DF, O'Neill, JP, and Whitfield, BL. (1981). The use of Chinese hamster ovary cells to quantify specific locus mutation and to determine mutagenicity of chemicals. Mutation Res. 86:193-214.
Myhr, BC and DiPaolo, JA. (1978). Mutagenesis of Chinese hamster cells in vitro by combination treatments with methyl methanosulfonate and N-acetoxy-2-acetylamino-fluorene. Cancer Res. 38: 2539-2543. - GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- CHO-K1-BH4 clone
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 from livers of aroclor-induced Sprague-Dawley rats
- Test concentrations with justification for top dose:
- 0.01, 0.02, 0.04, 0.06, 0.07, 0.08, 0.1, 0.13 µl/ml without metabolic activation
0.02, 0.04, 0.06, 0.08, 0.1, 0.13, 0.16, 0.2 µl/ml with metabolic activation - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO 10 µl/m
- True negative controls:
- other: not applicable
- Positive controls:
- yes
- Positive control substance:
- other: methylmethanesulfonate at 15 and 20 µl/ml, 5-bromo-2'-deoxyuridine at 50 µl/ml
- Remarks:
- without metabolic activation
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- 10 µl/ml DMSO
- True negative controls:
- other: not applicable
- Positive controls:
- yes
- Positive control substance:
- 3-methylcholanthrene
- Remarks:
- with metabolic activation Migrated to IUCLID6: 5 µl/ml
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in suspension
DURATION
- Exposure duration: 4 hrs
- Expression time (cells in growth medium): 7 days
- Fixation time (start of exposure up to fixation or harvest of cells): 14 days
SELECTION AGENT (mutation assays): 4 micrograms/ml 6-thioguanine - Evaluation criteria:
- Treatment groups were evaulated for relative survival, relative population growth, absolute cloning efficiency, and mutation frequency.
- Statistics:
- Kastenbaum, MA, Bowman, KO. (1970). Tables for determining the statistical significance of mutation frequencies. Mutat. Res. 9: 527-549.
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Cytotoxic at concentrations =>0.07 µl/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- There was no evidence of mutagenicity as compared to controls.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
The test material is not mutagenic, however, it is cytotoxic at concentrations of 0.07 µl/ml or above. - Executive summary:
A mammalian cell gene mutation assay was performed on Chinese hamster ovary cells to assess the mutagenicity of high flash aromatic naphtha to mammalian cells. The test was performed both with (0.02, 0.04, 0.06, 0.08, 0.1, 0.13, 0.16, 0.2 microliters/ml) and without (0.01, 0.02, 0.04, 0.06, 0.07, 0.08, 0.1, 0.13 microliters/ml) metabolic activation. Cells were then examined for mutation frequency. Only positive controls showed a significant increase in mutation frequency. No increase in mutation frequency was seen at any concentration either with or without metabolic activation. An analysis of relative population growth showed a reduction in population growth to 35.2% or greater of negative controls at exposures to concentrations or 0.07 microliters/ml or above. The test substance is therefore not mutagenic, however, it is cytotoxic at concentrations of 0.07 µl/ml or above.
- Endpoint:
- in vitro DNA damage and/or repair study
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1989
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Acceptable well-documented publication which meets basic scientific principles.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 479 (Genetic Toxicology: In Vitro Sister Chromatid Exchange Assay in Mammalian Cells)
- GLP compliance:
- yes
- Type of assay:
- sister chromatid exchange assay in mammalian cells
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 from livers of aroclor-induced Sprague-Dawley rats
- Test concentrations with justification for top dose:
- 2.0, 6.67, 20.0, 35.0, 50.1, 66.7 µg/ml without metabolic activation
0.667, 2.0, 6.67, 15.0, 20.0, 35.0, 50.1 µg/ml with metabolic activation - Vehicle / solvent:
- DMSO
- Untreated negative controls:
- yes
- Remarks:
- DMSO
- Negative solvent / vehicle controls:
- other: not applicable
- True negative controls:
- other: not applicable
- Positive controls:
- yes
- Positive control substance:
- other: MMC (unactivated cultures) and CP (metabolic activation)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in suspension
DURATION
- Exposure duration: 22.5 hrs (w/o activation); 2 hrs (w/ activation)
- Expression time (cells in growth medium): After 23 hour incubation with Brdu), metaphase cells were collected by mitotic shake off
Positive Controls: mitomycin C (MMC, unactivated cultures) and cyclophosphamide (CP, metabolic activation) - Evaluation criteria:
- Total sister chromatid exchange (SCE), SCE per chromosome, SCE per cell in the M2 stage of mitosis, percentage of cells in the M1, M1+, or M2 stages of cell cycle.
- Statistics:
- Evaluation of mutagenic responses was based on statistical comparisons of SCE frequencies in the C9 treated cultures with those in the negative (solvent) control cultures (Bancroft 1957, Hollander and Wolfe 1973).
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- cytotoxic at 90.2 µg/mL with metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- There were no increases in SCE at any concentration level tested in either the presence or absence of metabolic activation. Cell-cycle delay was not observed at concentration levels below 66.7 µg/mL (w/o activation) or below 50.1 µg/mL (w/ activation). The positive controls produced significant increases in the % SCE as compared to their respective negative controls.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
The test material did not cause sister chromatid exchange in CHO cells, however it is cytotoxic at the highest concentrations tested. - Executive summary:
The study examined the potential of high flash aromatic naphtha to cause sister chromatid exchange in Chinese hamster ovary (CHO) cells. CHO cells were exposed to test substance concentrations of 2.0, 6.67, 20.0, 35.0, 50.1, 66.7 micrograms/ml without metabolic activation, and 0.667, 2.0, 6.67, 15.0, 20.0, 35.0, 50.1 micrograms/ml with metabolic activation. The sister chromatid exchange frequency of cells in the treatment groups was then compared to the frequency in the control group. There were no increases in SCE at any concentration level tested in either the presence or absence of metabolic activation. The test material did not cause sister chromatid exchange in CHO cells, however it was cytotoxic at the highest concentrations tested.
Referenceopen allclose all
Mean revertant colonies (-S-9) - Experiment 1
Substance |
Dose level (µg/plate) |
TA98 |
TA100 |
TA1535 |
TA1537 |
TA102 |
Mean ± SD |
Mean ± SD |
Mean ± SD |
Mean ± SD |
Mean ± SD |
||
DMSO |
100 µl |
28 ± 6 |
109 ± 6 |
19 ± 5 |
12 ± 3 |
271 ± 18 |
SHELLSOL A |
0.533 |
26 ± 6 |
101 ± 10 |
14 ± 2 |
10 ± 8 |
251 ± 10 |
2.67 |
31 ± 1 |
108 ± 10 |
14 ± 2 |
9 ± 3 |
255 ± 15 |
|
13.3 |
26 ± 6 |
98 ± 6 |
17 ± 1 |
10 ± 1 |
277 ± 4 |
|
66.7 |
24 ± 5 |
91 ± 14 |
18 ± 6 |
9 ± 2 |
214 ± 2 |
|
333 (V+Ppn) |
21 ± 4 (V+Ppn) |
86 ± 0 (V+Ppn) |
13 ± 2 (V+Ppn) |
6 ± 2 (S+Ppn) |
184 ± 3 (S+Ppn) |
|
Positive Controls |
Compound |
2NF |
NaN3 |
NaN3 |
AAC |
GLU |
Dose Level |
5 µg |
2 µg |
2 µg |
50 µg |
25 µg |
|
Mean ± SD |
754 ± 130 |
509 ± 29 |
312 ± 27 |
376 ± 27 |
435 ± 28 |
Mean Revertant Colonies (+S-9) - Experiment 1
Substance |
Dose level (µg/plate) |
TA98 |
TA100 |
TA1535 |
TA1537 |
TA102 |
Mean ± SD |
Mean ± SD |
Mean ± SD |
Mean ± SD |
Mean ± SD |
||
DMSO |
100 µl |
32 ± 8 |
134 ± 9 |
17 ± 6 |
14 ± 5 |
318 ± 21 |
SHELLSOL A |
2.67 |
37 ± 5 |
143 ± 11 |
17 ± 7 |
11 ± 1 |
335 ± 45 |
8 |
41 ± 9 |
131 ± 7 |
21 ± 2 |
14 ± 3 |
315 ± 15 |
|
40 |
33 ± 6 |
133 ± 20 |
17 ± 3 |
15 ± 4 |
323 ± 12 |
|
200 |
34 ± 10 S |
125 ± 2 S |
23 ± 5 |
12 ± 2 |
275 ± 19 |
|
1000 |
31 ± 1 S |
112 ± 6 S |
15 ± 4 S |
9 ± 2 S |
252 ± 17 S |
|
Positive Controls |
Compound |
AAN |
AAN |
- |
- |
- |
Dose Level |
5 µg |
5 µg |
- |
- |
- |
|
Mean ± SD |
925 ± 121 |
1226 ± 104 |
- |
- |
- |
Mean Revertant Colonies (-S-9) - Experiment 2
Substance |
Dose level (µg/plate) |
TA98 |
TA100 |
TA1535 |
TA1537 |
TA102 |
Mean ± SD |
Mean ± SD |
Mean ± SD |
Mean ± SD |
Mean ± SD |
||
DMSO |
100 µl |
29 ± 7 |
117 ± 7 |
22 ± 5 |
5 ± 3 |
331 ± 19 |
SHELLSOL A |
31.25 |
26 ± 6 |
131 ± 11 |
20 ± 7 |
10 ± 2 |
304 ± 6 |
62.5 |
30 ± 6 |
113 ± 7 |
20 ± 4 |
8 ± 2 |
276 ± 9 |
|
125 |
27 ± 5 S |
113 ± 21 S |
18 ± 3 |
10 ± 2 |
278 ± 28 |
|
250 |
16 ± 1 S |
111 ± 19 S |
16 ± 1 S |
7 ± 2 S |
180 ± 6 S |
|
500 |
15 ± 2 M+Ppn+V |
100 ± 19 M+Ppn+S/V |
17 ± 6 M+S |
5 ± 2 S |
199 ± 11 S/V |
|
Positive Controls |
Compound |
2NF |
NaN3 |
NaN3 |
AAC |
GLU |
Dose Level |
5 µg |
2 µg |
2 µg |
50 µg |
25 µg |
|
Mean ± SD |
1004 ± 95 |
722 ± 15 |
438 ± 30 |
410 ± 112 |
595 ± 11 |
Mean Revertant Colonies (+S-9) Experiment 2
Substance |
Dose level (µg/plate) |
TA98 |
TA100 |
TA1535 |
TA1537 |
TA102 |
Mean ± SD |
Mean ± SD |
Mean ± SD |
Mean ± SD |
Mean ± SD |
||
DMSO |
100 µl |
38 ± 4 |
140 ± 7 |
18 ± 5 |
7 ± 3 |
360 ± 19 |
SHELLSOL A |
3.90625 |
- |
- |
22 ± 2 M |
- |
- |
7.8125 |
- |
- |
19 ± 2 |
10 ± 2 |
344 ± 13 |
|
12.5 |
36 ± 7 |
143 ± 9 |
- |
- |
- |
|
15.625 |
- |
- |
12 ± 4 M |
6 ± 3 |
323 ± 35 |
|
25 |
32 ± 9 |
133 ± 4 |
- |
- |
- |
|
31.25 |
- |
- |
17 ± 5 |
9 ± 1 |
287 ± 66 |
|
50 |
34 ± 9 |
134 ± 3 |
- |
- |
- |
|
62.5 |
- |
- |
17 ± 0 |
10 ± 1 |
307 ± 23 |
|
100 |
24 ± 3 S |
133 ± 4 S |
- |
- |
- |
|
125 |
- |
- |
17 ± 5 S |
4 ± 3 S |
242 ± 15 S |
|
200 |
26 ± 0 S |
131 ± 4 S |
- |
- |
- |
|
250 |
- |
- |
- |
5 ± 3 S |
209 ± 27 S |
|
Positive Controls |
Compound |
AAN |
AAN |
AAN |
AAN |
- |
Dose Level |
5 µg |
5 µg |
5 µg |
5 µg |
- |
|
Mean ± SD |
1109 ± 28 |
1045 ± 16 |
68 ± 3 |
63 ± 4 |
- |
S - Slight thinning of background lawn
M - Plate counted manually
Ppn - Precipitation observed
V - Very thin background lawn
AAN - 2 -aminoanthracene
2NF - 2 -nitrofluorene
NaN3 - sodium azide
AAC - 9 -aminoacridine
GLU - gluteraldehyde
Summary of other bacterial mutagenicity studies.
| End Point | Study Reference | ||||||||||||||||||||||||
| REACH requirement | IUCLID Section | Study Name | Data Waiving | Waiving Justification | Species | Study Result Type | Test Guideline/Qualifier | Test Guideline/Guideline | Test Guideline/Deviations | Reliability | Rational For Reliability | GLP Compliance | Test Materials/Identity | Study Result | Reference Type | Reference Author | Reference Year | Reference Title | Bibliographic Source | Testing Laboratory | Reference Report No. | Owner Company | Company Study No. | Report Date | Data access |
| 8.4.1 In vitro bacterial mutagenicity | 7.6.1 | The mutagenic potential of high flash aromatic naptha | Salmonella typhimurium | experimental result | According to | Ames, B.N., McCann, J., and Yamasaki, E. (1975). Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mutat. Res.31: 347-364. | No | 1 | Well-documented journal article. | No | Test material was specifically prepared to meet the composition requirements for High Flash Aromatic Naptha, Type 1 | not mutagenic | study report | Schreiner, CA, Edwards, DA, McKee, RH, Swanson, M, Wong, ZA, Schmitt, S, Beatty, P | 1989 | The mutagenic potential of high flash aromatic naptha | Cell Biology and Toxicology, Vol. 5, No. 2, 169-188 | yes | |||||||
CHO/HGPRT Forward Mutation Suspension Assay without Metabolic Activation
Mean Colony Number |
Relative Population Growth (%) |
Mutant Frequency in 10-6 units |
|
DMSO |
202.7 ± 7.6 |
111.0 |
1.0 |
DMSO |
190.0 ±17.8 |
89.0 |
2.2 |
5-bromo-2’-deoxyuridine |
161.3 ±11.2 |
114.1 |
14.0 |
Methyl methanesulfonate 15 µl/ml |
83.0 ±7.0 |
63.5 |
59.2 |
Methyl methanesulfonate 20 µl/ml |
41.0 ±7.2 |
38.3 |
59.4 |
0.01 µl/ml |
185.0 ±10.6 |
176.6 |
1.1 |
0.02 µl/ml |
204.7± 1.5 |
148.6 |
0.0 |
0.04 µl/ml |
204.3 ±2.5 |
147.5 |
1.9 |
0.06 µl/ml |
202.7± 20.5 |
107.5 |
0.6 |
0.07 µl/ml |
77.3 ±7.0 |
35.2 |
1.4 |
0.08 µl/ml |
5.7 ±1.5 |
10.7 |
0.9 |
0.1 µl/ml |
0.0± 0.0 |
ND |
ND |
0.13 µl/ml |
0.0 ±0.0 |
ND |
ND |
CHO/HGPRT Forward Mutation Suspension Assay with Metabolic Activation
Mean Colony Number |
Relative Population Growth (%) |
Mutant Frequency in 10-6 units |
|
DMSO |
203.7 ± 16.9 |
90.5 |
0.9 |
DMSO |
201.0 ± 12.5 |
109.5 |
4.4 |
3-methylcholanthene |
201.0 ± 7.8 |
77.1 |
161.3 |
0.02 µl/ml |
185.3 ± 3.5 |
119.7 |
2.6 |
0.04 µl/ml |
205.3 ± 21.1 |
111.7 |
3.4 |
0.06 µl/ml |
196.7 ± 22.0 |
110.0 |
3.4 |
0.07 µl/ml |
3.3 ± 1.5 |
4.0 |
1.3 |
0.08 µl/ml |
0.0 ± 0.0 |
ND |
ND |
0.1 µl/ml |
0.0 ± 0.0 |
ND |
ND |
0.13 µl/ml |
0.0 ± 0.0 |
ND |
ND |
0.16 µl/ml |
0.0 ± 0.0 |
ND |
ND |
0.2 µl/ml |
0.0 ± 0.0 |
ND |
ND |
Chromosome Aberrations in CHO Cells Exposed to High-Flash Aromatic Naphtha
| Dose | Cells | # Chromosomes | # SCE | SCE | SCE/Cell | |
| ug/mL | Scored | Chromosomes | (Mean +/- SE) | |||
| Assay #1: w/o metabolic activation | ||||||
| Negative Control | - | 50 | 1044 | 443 | 0.42 | 8.86 (.36) |
| Solvent Control | 11 | 50 | 1038 | 536 | 0.52 | 10.72 (.45) |
| Positive Control: MMC | 0.005 | 20 | 420 | 570 | 1.36 | 28.5 (1.13) |
| C9 Aromatics | 2 | 50 | 1037 | 530 | 0.51 | 10.6 (.43) |
| 6.67 | 50 | 1038 | 474 | 0.46 | 9.48 (.51) | |
| 20 | 50 | 1044 | 480 | 0.46 | 9.6 (.44) | |
| 66.67 | 50 | 1038 | 524 | 0.5 | 10.48 (.39) | |
| 200 | Toxic | - | - | - | - | |
| Assay #2: w/o metabolic activation | ||||||
| Negative Control | - | 50 | 1038 | 399 | 0.38 | 7.98 (.38) |
| Solvent Control | 11 | 50 | 1047 | 432 | 0.41 | 8.64 (.5) |
| Positive Control: MMC | 0.005 | 20 | 417 | 547 | 1.31 | 27.35 (1.49) |
| C9 Aromatics | 35 | 50 | 1043 | 428 | 0.41 | 8.56 (.49) |
| 50.1 | 50 | 1042 | 461 | 0.44 | 9.22 (.36) | |
| 66.7 | 50 | 1041 | 443 | 0.43 | 8.86 (.44) | |
| 90.1 | 50 | Toxic | - | - | - | |
| Dose | Cells | # Chromosomes | # SCE | SCE | SCE/Cell | |
| ug/mL | Scored | Chromosomes | (Mean +/- SE) | |||
| Assay #1: w/ metabolic activation | ||||||
| Negative Control | - | 50 | 1037 | 443 | 0.43 | 8.86 (.43) |
| Solvent Control | 11 | 50 | 1032 | 430 | 0.42 | 8.60 (.49) |
| Positive Control: MMC | 1.5 | 20 | 415 | 379 | 0.91 | 18.95 (1.20) |
| C9 Aromatics | 0.667 | 50 | 1038 | 449 | 0.43 | 8.98 (.34) |
| 2 | 50 | 1034 | 484 | 0.47 | 9.68 (.43) | |
| 6.67 | 50 | 1045 | 474 | 0.45 | 9.48 (.46) | |
| 20 | 50 | 1040 | 441 | 0.42 | 8.82 (.45) | |
| 66.67 | 50 | Toxic | - | - | - | |
| Assay #2: w/ metabolic activation | ||||||
| Negative Control | - | 50 | 1048 | 417 | 0.4 | 8.34 (.43) |
| Solvent Control | 11 | 50 | 1046 | 398 | 0.38 | 7.96 (.38) |
| Positive Control: MMC | 1.5 | 20 | 418 | 457 | 1.09 | 22.85 (.91) |
| C9 Aromatics | 15 | 50 | 1043 | 372 | 0.36 | 7.44 (.40) |
| 20 | 50 | 1048 | 444 | 0.42 | 8.88 (0.44) | |
| 35 | 50 | 1055 | 400 | 0.38 | 8.00 (.46) | |
| 50.1 | 50 | 1047 | 420 | 0.4 | 8.4 (.48) | |
| 66.7 | 50 | Toxic | - | - | - |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
Hydrocarbons, C9 Aromatics:
Genetic Toxicity in vivo - Mammalian Bone Marrow Chromosome Aberration Test (OECD TG 475): Negative
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:
- experimental study
- 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.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)
- GLP compliance:
- yes
- Type of assay:
- chromosome aberration assay
- Species:
- rat
- Strain:
- Sprague-Dawley
- Sex:
- male/female
- Route of administration:
- inhalation: vapour
- Details on exposure:
- - Exposure apparatus: 16 m glass and steel chambers.
- Method of holding animals in test chamber: cages
- Source and rate of air: Air was provided by a separate HVAC system.
- Method of conditioning air: Air was filtered for particulates and temperature and humidity controlled.
- System of generating particulates/aerosols: Test atmosphere was generated by heating nitrogen to 200°C by passing it through a 1 L stainless steel cylinder with a 1500 W band heater. The nitrogen then passed through a glass column 7.6 cm diameter and 30 cm long packed with glass beads. Test material was delivered by a metering pump into Teflon tubing, to the bottom of the column. The liquid test substance vaporized as it went up the column with the nitrogen. The vapor then went into the test chambers where dilution with the chamber ventilation air produced the desired concentrations.- Temperature, humidity, pressure in air chamber: Air flow rate, temperature and relative humidity were monitored every half-hour during exposure.
TEST ATMOSPHERE
- Brief description of analytical method used: Measurements made hourly using gas-phase IR.
- Samples taken from breathing zone: yes - Duration of treatment / exposure:
- 6 hours/day
- Frequency of treatment:
- 5 days
- Post exposure period:
- 6, 24, or 48 hours
- Remarks:
- Doses / Concentrations:
150 ppm
Basis:
nominal conc.
153 (9.6) ppm measured - Remarks:
- Doses / Concentrations:
500 ppm
Basis:
nominal conc.
471 (13.1) ppm measured concentration - Remarks:
- Doses / Concentrations:
1500 ppm
Basis:
nominal conc.
1540 (48) ppm measured concentration - No. of animals per sex per dose:
- 15 male/15 female
- Control animals:
- yes, concurrent no treatment
- Positive control(s):
- 10 animals of each sex were exposed to cyclophosphamide
- Route of administration: injected intraperitoneally
- Doses / concentrations: 40 mg/kg - Tissues and cell types examined:
- bone marrow
- Details of tissue and slide preparation:
- DETAILS OF SLIDE PREPARATION: 4 slides per rat were stained with Giemsa.
METHOD OF ANALYSIS: Slides were scanned for well resolved metaphase spreads. Fifty metaphases per animal were evaluated. - Evaluation criteria:
- total number of chromosome aberrations, frequency of aberrations per metaphase, percent metaphases with one or more aberrations, percent metaphases with two or more aberrations
- Statistics:
- Kruskal-Wallis multiple group comparison test followed by the Mann-Whitney U test
- Key result
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- reduced body weight gain in 1500 ppm group
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results: negative
The test substance was not clastogenic at levels up to and including 1500 ppm. - Executive summary:
A rat bone marrow cytogenicity study was performed to determine the clastogenicity of high flash aromatic naphtha. 15 male and 15 female rats were exposed via inhalation to 150, 500, or 1500 ppm of high flash aromatic naphtha for 6 hrs/day for 5 days. Rats were sacrificed at 6, 24, or 48 hrs after end of exposure, and the bone marrow then examined for chromosome and chromatid aberrations. There was no increase in the number of aberrations as compared to negative controls in any of the exposure groups. A significant increase in the number of aberrations was seen in the positive control group, therefore the test is valid. In conclusion, the test substance is not clastogenic.
Reference
Chromosome Aberrations in Sprague-Dawley Rats
Post-Exposure Interval |
Exposure Group |
Number |
Number of Spreads |
Number of Aberrations |
% Abrr. Per Metaphase |
% Metaphases > 1 Abrr. |
% Metaphases > 2 Abrr. |
6-hours |
Air |
8 |
400 |
0 |
0 |
0 |
0 |
150 ppm |
9 |
450 |
0 |
0 |
0 |
0 |
|
500 ppm |
10 |
487 |
0 |
0 |
0 |
0 |
|
1500 ppm |
9 |
450 |
0 |
0 |
0 |
0 |
|
24 hours |
Air |
9 |
450 |
1 |
0.2 |
0.2 |
0 |
150 ppm |
10 |
482 |
0 |
0 |
0 |
0 |
|
500 ppm |
10 |
500 |
0 |
0 |
0 |
0 |
|
1500 ppm |
10 |
500 |
1 |
0.2 |
0.2 |
0 |
|
Cyclo-phosphamide |
9 |
453 |
130 |
28.7 |
14.6 |
8.2 |
|
48 hours |
Air |
4 |
200 |
0 |
0 |
0 |
0 |
150 ppm |
4 |
200 |
0 |
0 |
0 |
0 |
|
500 ppm |
4 |
200 |
0 |
0 |
0 |
0 |
|
1500 ppm |
3 |
150 |
0 |
0 |
0 |
0 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Hydrocarbons, C9 Aromatics:
Hydrocarbons, C9 aromatics are not mutagenic using in vitro or in vivo genotoxicity assays. In bacterial reverse mutation tests, Hydrocarbons,C9 aromatics were not mutagenic in the presence or absence of metabolic activation. Likewise, there were no mutagenic effects reported in an in vitro mammalian gene mutation test (HGPRT forward mutation assay). No in vitro chromosomal effects were reported in a Chinese hamster ovary assay that examined the Hydrocarbons, C9 aromatics. The Hydrocarbons, C9 aromatics, were not clastogenic in an in vivo mammalian bone marrow chromosome aberration test. These data demonstrate that these substances are not categorizable genotoxins either in vitro or in vivo. Furthermore, no there evidence of hyperplastic responses or pre-neoplastic lesions in sub-chronic and chronic repeat-dose studies in either the Hydrocarbons, C9 aromatics. All studies were conducted in a manner similar or equivalent to currently established OECD guidelines. Hydrocarbons, C9 aromatics are a non-genotoxic agent and classification is not warranted.
Hydrocarbons, C10 Aromatics:
Hydrocarbons, C10 aromatics are not mutagenic using an in vitro genotoxicity assay. In bacterial reverse mutation tests, hydrocarbons, C10 aromatics were not mutagenic in the presence or absence of metabolic activation.
These data demonstrate that hydrocarbons, C9-C11, aromatics are not categorizable genotoxins either in vitro or in vivo. Furthermore, no there evidence of hyperplastic responses or pre-neoplastic lesions in sub-chronic and chronic repeat-dose studies in either the C9 aromatics or the C10 aromatics. All studies were conducted in a manner similar or equivalent to currently established OECD guidelines.
Justification for classification or non-classification
Based on the negative results from read across in vitro and in vivo genotoxicity assays, Hydrocarbons, C9, aromatics do not warrant classification as genotoxins under Regulation (EC) 1272/2008 on classification, labelling 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.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
