Registration Dossier
Registration Dossier
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EC number: 266-928-5 | CAS number: 67701-03-5 This substance is identified by SDA Substance Name: C16-C18 alkyl carboxylic acid and SDA Reporting Number: 19-005-00.
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Studies on genotoxicity are available for the following fatty acid
category members:
in-vitro:
- Gene mutation in bacteria:
CAS# 142-62-1, C6: Ames: negative (Banduhn 1991)
CAS# 124-07-2, C8: Ames RL4: negative (Gloxhuber and Wallat 1981)
CAS# 90990-08-2, C8-18: Ames RL4: negative (Wallat 1982)
CAS# 67701-06-8, C12-18: Ames RL4: negative (Sterzel and Broschard 1999)
CAS# 143-07-7, C12: Ames RL4: negative (Gloxhuber and Wallat 1981)
CAS# 112-85-6, C22: Ames RL4: negative (Gloxhuber and Wallat 1981)
CAS# 112-85-6, C22: Ames: negative (Nakajima 2002)
- Gene mutation in mammalian cells:
CAS# 334-48-5, C10: Mouse Lymphoma Assay in-vitro: negative (Trenz 2010)
- Cytogenicity in mammalian cells:
CAS# 112-85-6, C22: Chromosomal Aberration test in-vitro: negative
(Nakajima 2002)
in-vivo:
No data available.
All available data on genotoxicity showed that fatty acids are not
genotoxic.
Link to relevant study records
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- GLP - Guideline study, tested with the source substance CAS 112-85-6. In accordance to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- not applicable
- Species / strain / cell type:
- mammalian cell line, other: Chinese hamster lung (CHL) cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Eagle-MEM liquid medium
- Properly maintained: yes - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 from rat liver, induced with phenobarbital and 5,6-benzoflavone
- Test concentrations with justification for top dose:
- -S9 mix (short-term exposure): 0, 875, 1750, 3500 µg/mL
+S9 mix (short-term exposure): 0, 875, 1750, 3500 µg/mL
-S9 mix (24-hour continuous exposure): 0, 350, 700, 1400, 2800 µg/mL
-S9 mix (48-hour continuous exposure): 0, 288, 575, 1150, 2300 µg/mL - Vehicle / solvent:
- 1.0 % Carboxymethylcellulose sodium
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: continuous exposure: Mitomycin C (0.05 µg/mL for 24 hours and 0.025 µg/mL for 48 hours); short-term exposure, Cyclophosphamide (12.5 µg/mL)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: 3 days
- Exposure duration: 6 (short-term exposure), 24, 48 hours
STAIN (for cytogenetic assays): Giemsa
NUMBER OF CELLS EVALUATED: 200
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth - Evaluation criteria:
- The frequency of polyploid cells or cells with abnormal structure of each test group were determined according to the criteria of Ishidate.
- Species / strain:
- mammalian cell line, other: Chinese hamster lung (CHL) cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: insoluble in water, soluble in alcohol, ether, chloroform and acetone
- Precipitation: observed on the slide of continuous exposure high dose group
RANGE-FINDING/SCREENING STUDIES: see 'Any other information on material and method incl. tables'
COMPARISON WITH HISTORICAL CONTROL DATA: this test was valid, since the frequency of chromosomal aberration in positive control was within background data. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
The test substance did not induce structural chromosomal aberrations in the absence or presence of an exogenous metabolic activation system. - Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Study period:
- 09 Apr - 18 Apr 1991
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- GLP - Guideline study, tested with the source substance CAS 142-62-1. In accordance to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- TA 98: his D3052
TA 100: his G46
TA 1535: his G46
TA 1537: his C3076
TA 1538 his D3052
TA 1535, 1537 and TA 1538 contain rfa- and uvrB-; TA 98 and TA 100 contain rfa-, uvrB- and R+ - Species / strain / cell type:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix (Aroclor 1254-induced rat liver S9 fractions)
- Test concentrations with justification for top dose:
- First test: 8, 40, 200, 1000 and 5000 µg/ plate
Second test: 3.1, 12.5, 50, 200 and 800 µg/ plate - Vehicle / solvent:
- - Vehicle/solvent used: bidist. water
- Untreated negative controls:
- yes
- Remarks:
- suspension medium Tween 80/ bidist. water and untreated fresh cell suspension
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Buffer and Tween 80 / H2O
- True negative controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- other: see remarks
- Remarks:
- sodium azide: strains TA 100 and TA 1535; 9-aminoacridine: TA 1537; 4-nitro-o-phenylendiamine: TA 98 and TA1538 without S9 mix and 2-aminoanthracene in all strains with S9 mix
- Evaluation criteria:
- A combination of the following criteria was considered as a positive result:
- The plate background of non-reverted bacteria did not show any growth reduction versus the respective negative controls
- The spontaneous mutation rates of each tester strain per plate were within the characteristic spontaneous mutation range
- As a rule, the positive controls showed mutation rates exceeding the control values of TA 100 at least by the factor 2.0 and those of the other tester strains at least by the factor 3.0
- At more than one dose tested, the test substance caused at least a 2.0-fold increase in comparison with the negative controls in the tester strain TA 100. For the other tester strains, an increase in the mutation rate of more than 3.0 above the corresponding negative controls was considered positive. - Species / strain:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at the concentration of 800 µg / plate and higher
- 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
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Study period:
- 28 Jun - 23 Aug 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- GLP - Guideline study, tested with the source substance CAS 334-48-5. In accordance to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5300 - In vitro Mammalian Cell Gene Mutation Test
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian cell gene mutation tests using the thymidine kinase gene
- Target gene:
- Thymidine Kinase (TK)
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 1640 complete medium
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes - Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital (80 mg/kg bw) and beta-naphtoflavone (100 mg/kg bw)-induced rat liver S9 mix
- Test concentrations with justification for top dose:
- Pre-Test:
Experiment 1:
- with and without metabolic activation: 0.5, 2, 4, 6, 8, 10 mM
Experiment 2:
- without metabolic activation. 0.005, 0.05, 0.2, 0.7, 1.3, 2.0 mM
Main Test:
Experiment 1:
- with metabolic activation: 0.70, 0.82, 0.94, 1.06, 1.18, 1.30, 1.42, 1.54 mM
- without metabolic activation. 0.22, 0.46, 0.58, 0.70, 0.82, 0.94, 1.06, 1.18 mM
Experiment 2:
- with metabolic activation: 1.0, 1.12, 1.24, 1.36, 1.48, 1.60, 1.72, 1.84 mM
- without metabolic activation. 0.0005, 0.001, 0.002, 0.005, 0.01, 0.06, 0.18, 0.3 mM - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: RPMI cell culture medium
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- since medium was used as solvent, no further solvent control was necessary
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- without metabolic activation Migrated to IUCLID6: 200 and 500 µg/mL dissolved in medium
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- since medium was used as solvent, no further solvent control was necessary
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- without metabolic activation Migrated to IUCLID6: 10 µg/mL, dissolved in 0.9% NaCl
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- since medium was used as solvent, no further solvent control was necessary
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- with metabolic activation Migrated to IUCLID6: 3.5 µg/mL, dissolved in DMSO (1% final concentration in medium)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 h (short-term exposure, Experiment 1 with and without metabolic activation, Experiment 2 with metabolic activation) or 24 h (long-term exposure, Experiment 2 without metabolic activation)
- Expression time (cells in growth medium): 72 h (short-term exposure) or 48 h (long-term exposure)
- Selection time (if incubation with a selection agent): 11 - 14 days
SELECTION AGENT (mutation assays): 5 µg/mL trifluorothymidine (TFT )
NUMBER OF REPLICATIONS:
NUMBER OF CELLS EVALUATED: ca. 2000/plate
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth; cloning efficiency; mitotic index - Evaluation criteria:
- There are several criteria for a positive result:
- clear and dose-related increase in the mutant frequency
- biologically relevant response (at least a 2-fold increase of mutant frequencies related to the respective negative control values and higher than the historical range of negative controls) for at least one of the dose groups
- combined with a positive effect in the mutant frequency, an increased occurrence of small colonies (slow growth colonies) indicated by a low large/Small colonies ratio (1.5 times the ratio of clastogenic control MMS and/or B[a]P) is an indication for potential clastogenic effects and/or chromosomal aberrations.
The test substance is considered to be negative if there is no biologically relevant increase in the induction of mutant cells above concurrent control levels, at any dose level. - Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 1.54 mM with metabolic activation and at 1.18 mM without metabolic activation in experiment 1, respectively; at 1.84 mM with metabolic activation and at 0.30 mM without metabolic activation in experiment 2, respectively.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH:within the physiological range
- Effects of osmolality: within the physiological range
- Precipitation: in the pre-test with metabolic activation from concentrations of 4 mM and higher
RANGE-FINDING/SCREENING STUDIES:
All mutant values were found to be within the range of the historical control data of the test facility BSL Bioservice (about 51 to 170 mutants per 10^6 cells)
COMPARISON WITH HISTORICAL CONTROL DATA:
All mutant values were found to be within the range of the historical control data of the test facility BSL Bioservice (about 51 to 170 mutants per 10^6 cells) - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- other: Only short abstract available
- Principles of method if other than guideline:
- Ames test according to Mutation Research 31, 347 - 364 (1975)
- GLP compliance:
- not specified
- 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:
- not applicable
- Species / strain / cell type:
- S. typhimurium TA 1538
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254-induced rat liver S9 mix
- Test concentrations with justification for top dose:
- 4, 20, 100, 500 and 2500 µg / plate
- Vehicle / solvent:
- - Vehicle/solvent used: DMSO
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 4-Nitro-o-phenylene diamine
- Remarks:
- without metabolic activation
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- without metabolic activation
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 2-Amino-anthracene
- Remarks:
- with metabolic activation
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and 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
- 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
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- other: Only short abstract available
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- limited data; only tested up to 2500 µg/plate
- Principles of method if other than guideline:
- Ames test according to Mutation Research 31, 347 - 364 (1975)
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his
- Species / strain / cell type:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254-induced rat liver S-9mix
- Test concentrations with justification for top dose:
- 4, 20, 100, 500 and 2500 µg / plate
- Vehicle / solvent:
- - Vehicle/solvent used: Tween 80 / H2O bidist.
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 4-Nitro-o-phenylenediamine
- Remarks:
- without metabolic activation
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- without metabolic activation
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 2-Amino-anthracene
- Remarks:
- with metabolic activation
- Species / strain:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and 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
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- other: Only short abstract available
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- limited data; only tested up to 1250 µg/plate
- Principles of method if other than guideline:
- Ames test according to Mutation Research 31, 347 - 364 (1975)
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538
- 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 mix
- Test concentrations with justification for top dose:
- 4, 20, 100, 500, 1250 µg /plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Tween 80 / H20
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: without S9 mix: 4-Nitro-o-phenylene diamine and Sodium azide; with S9 mix: 2-Amino-anthracene
- Species / strain:
- S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and 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:
- not specified
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- GLP - Guideline study, tested with the source substance CAS 112-85-6. In accordance to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
- Qualifier:
- according to guideline
- Guideline:
- JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his, trp operon
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- E. coli WP2 uvr A
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix from livers of male Sprague-Dawley rats; induced with phenobarbital and 5,6.benzoflavone
- Test concentrations with justification for top dose:
- 156, 313, 625, 1250, 2500, 5000 µg/plate
- Vehicle / solvent:
- DMSO
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: -S9: 2-(2 Furyl)-3-(5-nitro-2-furyl)acrylamide (0.01 µg/plate); Sodium azide (0.5 µg/plate); Aminoacridine hydrochloride (80 µg/plate); +S9: 2-Aminoanthracene (1.0 µg/plate; TA100), (2 µg/plate; TA1535; TA1537), (10 µg/plate; WP2 uvrA), (0.5 µg/plate; TA9
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: preincubation
DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours
NUMBER OF REPLICATIONS: 2 times (3 plates/concentration/test)
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth
- Evaluation criteria:
- An increase twice the number of revertant colonies in test concentration plates compared to the solvent control plates and an observable dose-dependency is considered as a positive result.
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Additional information on results:
- The test substance did not induce gene mutations in the S. typhimurium and E. coli strains. No toxicity was observed up to a concentration of 5000 µg/plate, with or without metabolic activation.
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- other: Only short abstract available
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- limited data; only tested up to 2500 µg/plate
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- S. typhimurium TA 1538
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254-induce rat liver S-9 mix
- Test concentrations with justification for top dose:
- 4, 20, 100, 500, 2500 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water/Tween 80
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and 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:
- not specified
- 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:
- not specified
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- other: Only short abstract available
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- limited data; only 4 strains tested; only up to 2500µg/plate tested
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- S. typhimurium TA 1538
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor-induced rat liver S-9 mix
- Test concentrations with justification for top dose:
- 4, 20, 100, 500, 2500 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water and propylene glycol (Tween 80)
- Justification for choice of solvent/vehicle: substance is not soluble in water - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 4-nitro-o-phenyldiamine
- Remarks:
- without metabolic activation
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- without metabolic activation
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- with metabolic activation
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and 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
- 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
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results: negative
Referenceopen allclose all
Table 1: Results of growth inhibition test
Test item |
Concentration in µg/mL |
Survival in % |
Exposure period 24 h, without S9 mix |
||
1% CMC▪Na |
|
100 |
Test substance |
272 |
97.7 |
454 |
95.2 |
|
756 |
96.5 |
|
1260 |
83.1 |
|
2100 |
62.1 |
|
3500 |
37.8 |
|
Exposure period 48 h, without S9 mix |
||
1% CMC▪Na |
|
100 |
Test substance |
272 |
101.1 |
454 |
104.6 |
|
756 |
98.9 |
|
1260 |
89.7 |
|
2100 |
75.9 |
|
3500 |
1.3 |
|
Exposure period 6 h, without S9 mix |
||
1% CMC▪Na |
|
100 |
Test substance |
272 |
109.6 |
454 |
93.7 |
|
756 |
102.5 |
|
1260 |
110.7 |
|
2100 |
89.8 |
|
3500 |
95.1 |
|
Exposure period 6 h, with S9 mix |
||
1% CMC▪Na |
|
100 |
Test substance |
272 |
84.3 |
454 |
85.9 |
|
756 |
85.3 |
|
1260 |
86.5 |
|
2100 |
68.3 |
|
3500 |
76.3 |
|
Table 2: Results of chromosome aberration test
Test item |
Concentration |
Aberrant cells in % |
Polyploid cells in % |
||
|
in µg/mL |
with gaps |
without gaps |
||
Exposure period 24 h, without S9 mix |
|||||
1% CMC▪Na |
|
0.5 |
0.5 |
0.0 |
|
MMC |
0.05 |
56.0 |
52.5 |
0.5 |
|
Test substance |
350 |
1.0 |
1.0 |
0.5 |
|
700 |
1.0 |
0.5 |
0.5 |
||
1400 |
0.5 |
0.0 |
0.0 |
||
2800 |
Toxic |
||||
Exposure period 48 h, without S9 mix |
|||||
1% CMC▪Na |
|
0.0 |
0.0 |
0.5 |
|
MMC |
0.025 |
58.5 |
54.5 |
0.0 |
|
Test substance |
288 |
1.0 |
1.0 |
0.0 |
|
575 |
0.5 |
0.0 |
0.5 |
||
1150 |
2.0 |
1.5 |
0.0 |
||
2300 |
Toxic |
||||
Exposure period 6 h, without S9 mix |
|||||
1% CMC▪Na |
|
1.5 |
0.5 |
0.5 |
|
CP |
12.5 |
0.5 |
0.0 |
0.5 |
|
Test substance |
875 |
0.5 |
0.5 |
0.0 |
|
1750 |
3.0 |
2.5 |
0.0 |
||
3500 |
1.0 |
0.5 |
0.5 |
||
Exposure period 6 h, with S9 mix |
|||||
1% CMC▪Na |
|
1.5 |
1.0 |
0.0 |
|
CP |
12.5 |
63.5 |
61.5 |
0.0 |
|
Test substance |
875 |
2.5 |
1.5 |
1.0 |
|
1750 |
2.0 |
2.0 |
0.0 |
||
3500 |
2.0 |
2.0 |
0.0 |
||
CMC▪Na: carboxymethylcellulose sodium (solvent)
MMC: Mitomycin C; CP: Cyclophosphamide (positive controls)
Edenor C 6 98/100 did not induce any reverse mutations in the absence of rat liver enzymes (without S9-mix), nor did occur any induced reverse mutations in the presence of Aroclor 1254-induced S9-mix. Toxic effects were noted at the concentration of 800 µg / plate and higher.
In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test article did not induce point mutations by base-pair changes or frameshifts in the genome of the strains used.
Therefore, Edenor C 6 98/100 is not considered to be mutagenic in this Salmonella typhimurium reverse mutation assay.
Table 1. Test results of experiment 1 (plate incorporation)
With or without S9-Mix |
Test substance concentration (μg/plate) |
Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation) |
||||
Base-pair substitution type |
Frameshift type |
|||||
TA 100 |
TA1535 |
TA1538 |
TA98 |
TA1537 |
||
– |
Solvent: Tween 80/H2O |
71 ± 6 |
12 ± 3 |
10 ± 3 |
25 ± 4 |
6 ± 1 |
– |
8 |
78 ± 12 |
11 ± 4 |
9 ± 4 |
30 ± 2 |
9 ± 2 |
– |
40 |
74 ± 6 |
8 ± 4 |
9 ± 4 |
26 ± 10 |
5 ± 1 |
– |
200 |
69 ± 2 |
8 ± 2 |
12 ± 5 |
30 ± 3 |
8 ± 4 |
– |
1000 |
37 ± 8 r |
10 ± 3 |
2 ± 1 r |
11 ± 4 r |
2 ± 1 r |
– |
5000 |
I |
I; PP |
I |
I |
I |
Positive controls, –S9 |
Name |
SA |
SA |
4NPD |
4NPD |
9AA |
Concentrations (μg/plate) |
2 |
2 |
40 |
40 |
80 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
290 ± 40 |
477 ± 43 |
1517 ± 7 |
681 ± 38 |
610 ± 11 |
|
+ |
Solvent: Tween 80/H2O |
76 ± 5 |
12 ± 5 |
22 ± 3 |
48 ± 3 |
8 ± 2 |
+ |
8 |
91 ± 15 |
11 ± 5 |
16 ± 4 |
41 ± 4 |
8 ± 3 |
+ |
40 |
85 ± 7 |
10 ± 2 |
17 ± 1 |
34 ± 6 |
6 ± 1 |
+ |
200 |
72 ± 5 |
11 ± 5 |
17 ± 2 |
35 ± 4 |
7 ± 1 |
+ |
1000 |
56 ± 5 r |
9 ± 5 |
18 ± 3 |
12 ± 6 r |
4 ± 1 r |
+ |
5000 |
6 ± 5 |
I; PP |
I; PP |
I; PP |
I; PP |
Positive controls, +S9 |
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
Concentrations (μg/plate) |
5 |
2.5 |
5 |
5 |
2.5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
783 ± 41 |
164 ± 34 |
968 ± 97 |
881 ± 55 |
77 ± 21 |
|
I: Complete inhibition of the bacterial background lawn
r: Partial inhibition of the bacterial background lawn
PP: Pin-point colonies
9AA = 9-aminoacridine
SA: Sodium azide
4NPD: 4-Nitro-o-phenylenediamine
2AA = 2-Aminoanthracene
Table 2. Test results of experiment 2 (plate incorporation)
With or without S9-Mix |
Test substance concentration (μg/plate) |
Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation) |
||||
Base-pair substitution type |
Frameshift type |
|||||
TA 100 |
TA1535 |
TA1538 |
TA98 |
TA1537 |
||
– |
Solvent: Tween 80/H2O |
72 ± 7 |
10 ± 4 |
8 ± 2 |
31 ± 5 |
8 ± 2 |
– |
3.1 |
65 ± 1 |
8 ± 1 |
7 ± 2 |
30 ± 4 |
8 ± 5 |
– |
12.5 |
72 ± 4 |
15 ± 6 |
8 ± 1 |
32 ± 7 |
7 ± 3 |
– |
50 |
67 ± 11 |
8 ± 3 |
7 ± 6 |
33 ± 1 |
7 ± 1 |
– |
200 |
73 ± 9 |
10 ± 2 |
8 ± 5 |
29 ± 6 |
5 ± 3 |
– |
800 |
45 ± 12 r |
8 ± 5 r |
5 ± 2 r |
23 ± 4 r |
4 ± 2 r |
Positive controls, –S9 |
Name |
SA |
SA |
4NPD |
4NPD |
9AA |
Concentrations (μg/plate) |
2 |
2 |
40 |
40 |
80 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
314 ± 22 |
499 ± 21 |
1659 ± 63 |
629 ± 10 |
739 ± 33 |
|
+ |
Solvent: Tween 80/H2O |
79 ± 3 |
13 ± 2 |
16 ± 5 |
41 ± 7 |
12 ± 1 |
+ |
3.1 |
78 ± 7 |
10 ± 1 |
18 ± 3 |
38 ± 9 |
10 ± 1 |
+ |
12.5 |
70 ± 7 |
7 ± 1 |
15 ± 5 |
36 ± 8 |
10 ± 1 |
+ |
50 |
82 ± 15 |
12 ± 4 |
19 ± 2 |
33 ± 1 |
8 ± 4 |
+ |
200 |
78 ± 7 |
12 ± 3 |
20 ± 2 |
38 ± 8 |
10 ± 3 |
+ |
800 |
60 ± 13 |
11 ± 3 r |
11 ± 1 r |
26 ± 5 |
5 ± 2 r |
Positive controls, +S9 |
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
Concentrations (μg/plate) |
5 |
2.5 |
5 |
5 |
2.5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
862 ± 112 |
215 ± 2 |
1300 ± 212 |
1000 ± 46 |
88 ± 14 |
|
I: Complete inhibition of the bacterial background lawn
r: Partial inhibition of the bacterial background lawn
PP: Pin-point colonies
9AA = 9-aminoacridine
SA = Sodium azide
4NPD = 4-Nitro-o-phenylenediamine
2AA = 2-Aminoanthracene
Colony sizing was performed for the highest concentrations of the test item and for the negative and positive controls. A mutation frequency above 2 in combination with an increased occurrence of small colonies (defined by slow growth and/or morphological alteration of the cell clone), indicated by a low large/small colony ratio (ratio of the clastogenic controls MMS and/or B[a]P with a coefficient of 1.5), is an indication for potential clastogenic effects and/or chromosomal aberrations.
Although in experiment 1 with metabolic activation an increased number of small colonies was noted at doses of 1.30 mM and 1.42 mM (26 and 31small colonies, respectively, compared to 11 and 9 at control) all dose groups were considered as not clastogenic since no mutagenicity was found at these doses.
All other dose groups in the other experiments were also found not to be clastogenic, respectively.
Table 1: Experiment I - 4 h exposure - With Metabolic Activation
Concentration |
Cloning efficiency [%] |
Relative Total Growth [%] |
Mutants per 1E+06 surviving cells |
Mutation factor |
Colony Sizing Quotient Large/Small |
0 |
100 |
100 |
86.77 |
1 |
3.66 |
0.7 |
96.63 |
95.51 |
95.86 |
1.10 |
-- |
0.82 |
104.12 |
92.71 |
88.56 |
1.02 |
-- |
0.94 |
109.36 |
95.60 |
76.43 |
0.88 |
-- |
1.06 |
110.11 |
78.73 |
78.58 |
0.91 |
-- |
1.18 |
109.36 |
60.06 |
86.55 |
1.00 |
-- |
1.30 |
116.10 |
40.14 |
100.88 |
1.16 |
1.77 |
1.42 |
112.36 |
31.61 |
121.24 |
1.40 |
1.55 |
1.54 |
96.63 |
9.84 |
139.92 |
1.61 |
2.78 |
B[a]P, 3.5 µg/mL |
99.63 |
69.03 |
623.89 |
7.19 |
1.24 |
B[a]P Benzo[a]pyrene
Table 2: Experiment I - 4 h exposure - Without Metabolic Activation
Concentration |
Cloning efficiency [%] |
Relative Total Growth [%] |
Mutants per 1E+06 surviving cells |
Mutation factor |
Colony Sizing Quotient Large/Small |
0 |
100 |
100 |
79.39 |
1 |
2.75 |
0.22 |
100.33 |
90.09 |
78.53 |
0.99 |
-- |
0.46 |
86.38 |
79.76 |
124.63 |
1.57 |
-- |
0.58 |
91.03 |
79.70 |
77.88 |
0.98 |
-- |
0.70 |
98.34 |
89.53 |
70.89 |
0.89 |
-- |
0.82 |
98.34 |
71.30 |
69.28 |
0.87 |
-- |
0.94 |
95.02 |
64.50 |
62.74 |
0.79 |
1.60 |
1.06 |
99.00 |
47.66 |
74.59 |
0.94 |
3.17 |
1.18 |
91.69 |
11.04 |
97.49 |
1.23 |
1.12 |
EMS, 500 µg/mL |
86.38 |
62.27 |
1337.77 |
16.85 |
-- |
MMS, 10 µg/mL |
85.71 |
66.62 |
841.03 |
10.59 |
0.69 |
EMS Ethyl methane sulphonate
MMS Methyl methane sulphonate
Table 3: Experiment II - 4 h Exposure - With Metabolic Activation
Concentration |
Cloning efficiency [%] |
Relative Total Growth [%] |
Mutants per 1E+06 surviving cells |
Mutation factor |
Colony Sizing Quotient Large/Small |
0 |
100 |
100 |
82.64 |
1.00 |
2.07 |
1 |
96.97 |
85.91 |
82.23 |
1.00 |
-- |
1.12 |
101.68 |
89.08 |
61.54 |
0.74 |
-- |
1.24 |
98.99 |
86.20 |
86.92 |
1.05 |
-- |
1.36 |
99.66 |
83.65 |
75.75 |
0.92 |
-- |
1.48 |
90.91 |
64.04 |
118.05 |
1.43 |
-- |
1.60 |
96.07 |
35.51 |
70.23 |
0.85 |
2.38 |
1.72 |
91.58 |
38.25 |
84.77 |
1.03 |
2.13 |
1.84 |
98.99 |
19.98 |
98.81 |
1.20 |
4.73 |
B[a]P, 3.5 µg/mL |
86.20 |
60.39 |
829.02 |
10.03 |
0.89 |
B[a]P Benzo[a]pyrene
Table 4: Experiment II - 24 h exposure - Without Metabolic Activation
Concentration |
Cloning efficiency [%] |
Relative Total Growth [%] |
Mutants per 1E+06 surviving cells |
Mutation factor |
Colony Sizing Quotient Large/Small |
0 |
100 |
100 |
104.66 |
1 |
2.61 |
0.0005 |
95.24 |
94.64 |
76.34 |
0.73 |
-- |
0.001 |
101.36 |
103.58 |
68.22 |
0.65 |
-- |
0.002 |
94.56 |
95.29 |
66.56 |
0.64 |
-- |
0.005 |
101.36 |
105.73 |
52.63 |
0.50 |
-- |
0.01 |
102.04 |
98.98 |
64.06 |
0.61 |
-- |
0.06 |
102.72 |
96.75 |
61.54 |
0.59 |
3.30 |
0.18 |
96.60 |
55.56 |
91.91 |
0.88 |
2.24 |
0.30 |
91.16 |
19.77 |
99.26 |
0.95 |
1.94 |
EMS, 200 µg/mL |
70.75 |
34.84 |
2516.55 |
24.05 |
-- |
MMS, 10 µg/mL |
59.18 |
27.33 |
2625.00 |
25.08 |
0.81 |
EMS Ethyl methane sulphonate
MMS Methyl methane sulphonate
Edenor C 12 98/100% did not induce reverse mutations in the absence or presence of S9 mix in the tester strains Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538.
Edenor C 8 98/100% did not induce reverse mutations in the absence or presence of S-9 mix in the tester strains Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538.
Edenor C 22 R did not induce reverse mutations in the absence or presence of S9 mix in the tester strains Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538.
Edenor C 22 R did not show mutagenic activity in vitro.
Table 1: Test Results of Experiment 1
EXPERIMENT 1 (Preincubation Test) |
|||||
S9-Mix |
Without |
||||
Test item (µg/plate) |
TA 100 |
TA 1535 |
E. coli WP2 uvrA |
TA 98 |
TA 1537 |
DMSO |
95 ± 2 |
15 ± 1 |
25 ± 2 |
23 ± 3 |
9 ± 2 |
156* |
94 ± 3 |
11 ± 1 |
27 ± 3 |
21 ± 2 |
8 ±3 |
313* |
94 ± 4 |
12 ± 2 |
24 ± 1 |
21 ± 4 |
10 ± 2 |
625* |
89 ± 2 |
14 ± 3 |
26 ± 1 |
18 ± 2 |
8 ± 1 |
1250* |
82 ± 4 |
13 ± 1 |
25 ± 3 |
21 ± 3 |
8 ± 3 |
2500* |
80 ± 2 |
11 ± 1 |
26 ± 1 |
17 ± 3 |
10 ± 4 |
5000* |
78 ± 2 |
9 ± 2 |
28 ± 1 |
14 ± 4 |
10 ± 3 |
AF-2 |
687 ± 12 |
- |
173 ± 10 |
460 ± 27 |
- |
NaN3 |
- |
422 ± 7 |
- |
- |
- |
ACR |
- |
- |
- |
- |
491 ± 28 |
S9-Mix |
With |
||||
Test item (µg/plate) |
TA 100 |
TA 1535 |
E. coli WP2 uvrA |
TA 98 |
TA 1537 |
DMSO |
112 ± 4 |
13 ± 1 |
31 ± 2 |
33 ± 2 |
7 ± 1 |
156* |
103 ± 4 |
12 ± 3 |
27 ± 3 |
30 ± 3 |
10 ± 3 |
313* |
106 ± 10 |
10 ± 4 |
26 ± 1 |
34 ± 4 |
11 ± 3 |
625* |
97 ± 3 |
12 ± 2 |
27 ± 1 |
29 ± 4 |
10 ± 3 |
1250* |
97 ± 5 |
12 ± 4 |
28 ± 5 |
30 ± 2 |
10 ± 1 |
2500* |
98 ± 3 |
12 ± 3 |
27 ± 3 |
30 ± 2 |
10 ± 3 |
5000* |
99 ± 9 |
9 ± 2 |
28 ± 2 |
29 ± 1 |
9 ± 1 |
2-AA |
723 ± 12 |
383 ± 5 |
516 ± 5 |
366 ± 19 |
172 ± 8 |
* precipitation occurred at the end of the exposure
AF-2: 2-(2 furyl)-3-(5-nitro-2-furyl)acrylamide (0.01 µg/plate for TA 100 and E. coli WP2 uvrA; 0.1 µg/plate for TA 98)
NaN3: sodium azide (0.5 µg/plate)
ACR: 9-aminoacridine hydrochloride (80 µg/plate)
2-AA: 2-aminoanthracene (1 µg/plate for TA 100; 2 µg/plate for TA 1535 and TA 1537; 10 µg/plate for E. coli WP2 uvrA; 0.5 µg/plate for TA 98)
Table 2: Test Results of Experiment 2
EXPERIMENT 2 (Preincubation Test) |
|||||
S9-Mix |
Without |
||||
Test item (µg/plate) |
TA 100 |
TA 1535 |
E. coli WP2 uvrA |
TA 98 |
TA 1537 |
DMSO |
95 ± 2 |
16 ± 2 |
26 ± 3 |
23 ± 2 |
5 ± 1 |
156* |
94 ± 3 |
14 ± 1 |
25 ± 1 |
23 ± 3 |
6 ± 2 |
313* |
95 ± 3 |
14 ± 3 |
26 ± 2 |
23 ± 4 |
6 ± 2 |
625* |
90 ± 0 |
17 ± 1 |
24 ± 3 |
22 ± 2 |
7 ± 0 |
1250* |
89 ± 5 |
15 ± 1 |
25 ± 3 |
21 ± 2 |
5 ± 2 |
2500* |
85 ± 5 |
15 ± 2 |
23 ± 1 |
20 ± 1 |
7 ± 1 |
5000* |
88 ± 3 |
15 ± 3 |
22 ± 2 |
20 ± 1 |
4 ± 1 |
AF-2 |
734 ± 14 |
- |
128 ± 6 |
439 ± 7 |
- |
NaN3 |
- |
433 ± 3 |
- |
- |
- |
ACR |
- |
- |
- |
- |
503 ± 5 |
S9-Mix |
With |
||||
Test item (µg/plate) |
TA 100 |
TA 1535 |
E. coli WP2 uvrA |
TA 98 |
TA 1537 |
DMSO |
107 ± 3 |
15 ± 2 |
29 ± 2 |
26 ± 2 |
14 ± 1 |
156* |
100 ± 2 |
15 ± 1 |
28 ± 1 |
29 ± 2 |
14 ± 2 |
313* |
99 ± 4 |
13 ± 1 |
29 ± 3 |
30 ± 8 |
14 ± 1 |
625* |
103 ± 3 |
14 ± 2 |
27 ± 2 |
30 ± 4 |
15 ± 2 |
1250* |
101 ± 4 |
12 ± 2 |
28 ± 5 |
28 ± 2 |
13 ± 1 |
2500* |
98 ± 2 |
13 ± 2 |
26 ± 2 |
26 ± 1 |
11 ± 1 |
5000* |
96 ± 5 |
10 ± 1 |
26 ± 2 |
24 ± 1 |
12 ± 2 |
2-AA |
678 ± 17 |
393 ± 5 |
521 ± 22 |
423 ± 31 |
173 ± 13 |
* precipitation occurred at the end of the exposure
AF-2: 2-(2 furyl)-3-(5-nitro-2-furyl)acrylamide (0.01 µg/plate for TA 100 and E. coli WP2 uvrA; 0.1 µg/plate for TA 98)
NaN3: sodium azide (0.5 µg/plate)
ACR: 9-aminoacridine hydrochloride (80 µg/plate)
2-AA: 2-aminoanthracene (1 µg/plate for TA 100; 2 µg/plate for TA 1535 and TA 1537; 10 µg/plate for E. coli WP2 uvrA; 0.5 µg/plate for TA 98)
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Fatty acids are found in all living organisms fulfilling fundamental physiological functions within the body. Based on this role within the body, no potential of fatty acids for genotoxicity is expected as it could be demonstrated in-vitro with fatty acids C6, C8, C10, C12, and C22, as well as with fatty acids mixtures C8-18 and C12-18.
Two of the available studies on gene mutation in bacteria were conducted according to OECD guideline 471 and under GLP conditions with hexanoic acid and docosanoic acid, respectively (C6: Banduhn, 1991; C22: Nakajiama, 2002). In both studies bacteria strains S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 were tested with the fatty acids at concentrations up to 5000 µg/plate with and without metabolic activation by rat liver S9-mix. C22 fatty acid was additionally tested with bacteria strain E. coli WP2 uvr A to detect DNA-cross linking. The test substances did not induce gene mutations in the S. typhimurium and E. coli strains. No toxicity was observed up to a concentration of 5000 µg/plate, with or without metabolic activation. The other available studies were insufficient for assessment due to limited documentation (C8: Gloxhuber and Wallat 1981; C8-18: Wallat 1982; C12-18: Sterzel and Broschard 1999; C12: Gloxhuber and Wallat 1981; C22: Gloxhuber and Wallat 1981). However, according to the authors, all these studies with fatty acids of different chain lengths gave negative results showing that fatty acids do not induce gene mutation in bacteria.
Regarding gene mutation in mammalian cells an in vitro mouse lymphoma assay was performed with decanoic acid under GLP according to OECD guideline 476 (Trenz, 2010). In two experiments, mouse lymphoma L5178Y cells were treated with decanoic acid at concentrations up to 1.54 mM with metabolic activation (phenobarbital and beta-naphtoflavone-induced rat liver S9-mix) and up to 1.18 mM without metabolic activation, respectively. Although cytotoxicity was observed , all mutant values were found to be within the range of the historical control data of the test facility, so that decanoic acid was not found to be mutagenic. In addition, colony sizing was performed for the highest concentrations used to detect potential clastogenic effects and/or chromosomal aberrations. As result, decanoic acid was not found to be clastogenic at all dose groups tested.
An in vitro mammalian chromosome aberration test was conducted with C22 fatty acid (docosanoic acid) in accordance with GLP and OECD guideline 473 and Japanese Guidelines for Screening Mutagenicity Testing of Chemicals (Nakajima, 2002). Properly maintained Chinese hamster lung (CHL) cells were treated with docosanoic acid dissolved in 1% carboxymethylcellulose sodium at concentrations of 875, 1750 and 3500 µg/mL with and without metabolic activation by S9 from Phenobarbital- and 5,6-benzoflavone-induced rat liver for 6 hours. In addition, the cells were incubated with 350, 700, 1400, 2800 µg/mL without metabolic activation for 24 hours and with 288, 575, 1150 and 2300 µg/mL without metabolic activation for 48 hours. No increase in chromosomal aberrations was found at all concentration tested.
Taking all the results together and based on a weight of evidence approach, the negative results for genetic toxicity, which were obtained with different members of the category do not provide any evidence that fatty acids are genotoxic.
Short description of key information:
Studies on genotoxicity are available for the following fatty acid
category members:
in-vitro:
- Gene mutation in bacteria:
CAS# 142-62-1, C6: Ames: negative (Banduhn 1991)
CAS# 124-07-2, C8: Ames RL4: negative (Gloxhuber and Wallat 1981)
CAS# 90990-08-2, C8-18: Ames RL4: negative (Wallat 1982)
CAS# 67701-06-8, C12-18: Ames RL4: negative (Sterzel and Broschard 1999)
CAS# 143-07-7, C12: Ames RL4: negative (Gloxhuber and Wallat 1981)
CAS# 112-85-6, C22: Ames RL4: negative (Gloxhuber and Wallat 1981)
CAS# 112-85-6, C22: Ames: negative (Nakajima 2002)
- Gene mutation in mammalian cells:
CAS# 334-48-5, C10: Mouse Lymphoma Assay in-vitro: negative (Trenz 2010)
- Cytogenicity in mammalian cells:
CAS# 112-85-6, C22: Chromosomal Aberration test in-vitro: negative
(Nakajima 2002)
in-vivo:
No data available.
All available data on genotoxicity showed that fatty acids are not
genotoxic.
Endpoint Conclusion: No adverse effect observed (negative)
Justification for classification or non-classification
According to CLP (1272/2008/EC) classification criteria for genetic toxicity, fatty acids do not fulfill the criteria for classification and thus a non-classification is warranted for this endpoint.
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