Registration Dossier
Registration Dossier
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EC number: 294-304-2 | CAS number: 91697-43-7
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 06 Mar - 09 May 1990
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions (incomplete strain selection; E. coli is missing).
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- incomplete strain selection; E. coli is missing
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his operon
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Standard I nutrient broth for microbiology (Merck art. no. 7882)
- Species / strain / cell type:
- S. typhimurium TA 1538
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Standard I nutrient broth for microbiology (Merck art. no. 7882)
- Metabolic activation:
- with and without
- Metabolic activation system:
- cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254.
- Test concentrations with justification for top dose:
- 1st and 2nd experiment: 8, 40, 200, 100 and 5000 µg/plate with and without metabolic activation
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Tween 80 / bidist. water
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- Migrated to IUCLID6: 2 µg/plate, TA100 and TA1535 with and without S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- Migrated to IUCLID6: 80 µg/plate, TA 1537 with and without S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 40 µg/plate 4-nitro-o-phenylendiamine, TA 98 and TA 1538 with and without S9
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-amino-anthracene: 2.5 µg/plate in TA1535an d TA1537; 5 µg/plate in TA100, TA1538 and TA98 with and without S9
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 hours
NUMBER OF REPLICATIONS: Two independent experiments were performed each in triplicates.
DETERMINATION OF CYTOTOXICITY
- Method: Reduction in the number of revertant colonies, background lawn - Evaluation criteria:
- A combination of the following criteria was considered as a positive result:
1) The plate background of non-reverted bacteria did not show any growth reduction versus the respective negative controls.
2) The spontaneous mutation rates of each tester strain per plate were within the characteristic spontaneous mutation range.
3) 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.
4) 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.
Reproducibility:
If the test substance induces reverse mutations in only one test, and if this effect cannot be reproduced in one or several repeated tests, the initially positive test data will lose their significance. - Statistics:
- No data
- 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
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1538
- 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
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative - Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Study period:
- 04 Nov - 02 Dec 2004
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- GLP guideline study, tested with the source substance CAS 163961-32-8. According 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
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- The Department of Health of the Government of the united Kingdom
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Duplicate cultures of human lymphocytes were evaluated for chromosome aberrations at up to three dose levels.
- Species / strain / cell type:
- primary culture, other: human lymphocytes
- Details on mammalian cell type (if applicable):
- For each experiment, whole blood was drawn from the peripheral circulation of a volunteer who had previously been screened for suitability. The cell cycle length of 17 h was determined by BrdU incorporation.
- Metabolic activation:
- with and without
- Metabolic activation system:
- PB/βNF induced S9-Mix from male Spraque-Dawley rats
- Test concentrations with justification for top dose:
- 1. Experiment:
24 h continous exposure without S9: 312.5; 468.75; 625 µg/mL
4 h exposure to test material with S9 mix followe by 20 h culture in treatment free media: 625; 1250 and 2500 µg/mL
2. Experiment
4 h exposure to test material without S9 mix followe by 20 h culture in treatment free media: 625; 1250 and 2500 µg/mL
4 h exposure to test material with S9 mix followe by 20 h culture in treatment free media: 625; 1250 and 2500 µg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: arachis oil
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Mitomycin C (MMC, 0.2 + 0.4µg/mL; -S9); Cyclophosphamide (CP, 7,5µg/mL; +S9)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium (Eagle´s minimal essential medium with HEPES buffer supplemented with L-glutamine, Pen/Strep, amphotericin B and 15% FCS
DURATION
- Exposure duration: Experiment I: 4 hours with S9 mix, or 24 hours without S9 mix; Experiment II: 4 hours with and without S9 mix
- Expression time (cells in growth medium): 20 hours after 4 hour exposure in both experiments
NUMBER OF REPLICATIONS: 2
SPINDLE INHIBITOR (cytogenetic assays): Colcemid (0.1 µg/mL)
STAIN (for cytogenetic assays): 5% Giemsa for 5 min
NUMBER OF CELLS EVALUATED: Where possible the first 100 consecutive well-spread metaphases from each culture were counted, where there were approximately 50% of cells with aberrations, slide evaluation was terminated at 50 cells. If the cell had 44-48 chromosomes, any gaps, breaks or reÍurangements were noted according to the simplified system of Savage (1976) recommended in the 1983 UKEMS guidelines for mutagenicrty testing.
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index: A total of 2000 or 1000 lymphocyte cell nuclei were counted and the number of cells in metaphase recorded and expressed as the mitotic index and as a percentage of the vehicle control value. - Evaluation criteria:
- A positive response was recorded for a particular treatment if the % cells with aberrations, excluding gaps, markedly exceeded that seen in the concurrent control, either with or without a clear dose-relationship. For modest increases in aberration frequency a dose response relationship is generally required and appropriate statistical tests may be applied in order to record a positive response.
- Species / strain:
- primary culture, other: human
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- max. 38% inhibition at 468,75 mg/mL and 24 h incubation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- primary culture, other: human
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Remarks:
- max. 11% (1st experiment) and 22% (2nd experiment) inhibition at 1250 mg/mL
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- RANGE-FINDING/SCREENING STUDIES: Dosing was based on that utilised in a previous study on a different batch of the test material (produced by a different manufacturing method) and included a dose level that induced some mitotic inhibition.
2500 µg/mL was chosen as maximum dose due to limited solubility. - Remarks on result:
- other: strain/cell type: lymphocytes
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative non-clastogenic to human lymphocytes in vitro
The numbers of chromosome aberrations and polyploid cells were not increased at any dose level in comparison to the control. The test substance is non-clastogenic in mammalian cells in vitro. - Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- migrated information: read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Study period:
- 05 Jan - 05 Mar 2007
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: GLP guideline study with acceptable restrictions (analytical purity of test substance not specified).
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- yes
- Remarks:
- analytical purity of test substance not specified
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- The Department of Health of the Government of the United Kingdom
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- thymidine kinase
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - details on cell strain: L5178Y +/- 3.7.2c mouse lymphoma cells
- Type and identity of media: RPMI 1640 medium with Glutamax-l and HEPES buffer
- Properly maintained: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- S9-Mix
- Test concentrations with justification for top dose:
- Experiment I: 4 hour treatment with and without S9 mix - 156.25, 312.5, 625, 1250, 2500, 5000 µg/mL
Experiment II: 24 hour treatment without and 4 hour treatment with S9 mix - 156.25, 312.5, 625, 1250, 2500, 5000 µg/mL
Experiment III: 4 hour treatment with S9 mix - 39.06, 78.13, 117.2, 156.25, 234.38, 312.5, 468.75, 625, 937.5, 1250 µg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Acetone
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Acetone
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: ethylmethanesulphonate (EMS, 150 + 400 µg/mL without S9); Cyclophosphamide (CP, 2 µg/mL with S9 mix)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 h with and without metabolic activation increased to 24 h in the second experiment (without metabolic activation)
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): 10-14 days
SELECTION AGENT (mutation assays): 5-fluorothymidine
NUMBER OF REPLICATIONS: 2
DETERMINATION OF CYTOTOXICITY
- Cell counts, viability after 2 days: the cell counts obtained immediately post treatment and over the 2-day expression period were used to calculate the % relative suspension growth that gives an indication of post treatment toxicity
OTHER:
The numbers of small and large colonies seen in the TFT mutation plates were also recorded - Evaluation criteria:
- - The vehicle control values have to be in range with historical data.
- Positive control chemicals should induce at least three to five fold increases in mutant frequency greater than the corresponding vehicle control.
- For a test material to demonstrate a mutagenic response it must produce a statistically significant increase in the induced mutant frequency (IMF) over the concurrent vehicle mutant frequency value. The IMF must exceed the Global Evaluation Factor (GEF).
- when a test material induces modest reproducible increases in the mutation frequencies that do not exceed the GEF value then scientific judgement will be applied. If the reproducible responses are significantly dose-related and include increases in the absolute numbers of mutant colonies then they may be considered to be toxicologically significant. - Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
A cloudy precipitate of the test material was observed at and above 78.13 µg/mL forming a greasy/oily precipitate at and above 1250 µg/mL.
RANGE-FINDING/SCREENING STUDIES:
In the 4-hour exposures, both in the absence and presence of metabolic activation (S9), there appeared to be variable exposure of the test material to the cells as indicated by the reductions in the Relative Suspension Growth (%RSG) at certain dose levels. This was considered to be due to the nature of the test material formulations. In the 24-hour exposure in the absence of S9 there was a greater reduction in %RSG values forming a plateau effect down the dose range. A cloudy precipitate of the test material was observed at and above 78.13 pglml forming a greasy/oily precipitate at and above 1250 pglml. Due to the variable levels of toxicity observed the maximum recommended dose of 5000 pgiml was investigated in Experiments I and II.
COMPARISON WITH HISTORICAL CONTROL DATA:
All results were within the ranges of historical control data.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
The dose range used in the preliminary toxicity test was 19.53 to 5000 µg/mL.
Due to the variable levels of toxicity observed the maximum recommended dose of 5000 µg/mL was investigated in experiments 1 and 2. - Conclusions:
- Interpretation of results (migrated information):
negative not mutagenic in mouse lymphoma cells
The test material did not induce any toxicologically significant increases in the mutant frequency at the TK +/- locus in L5l78Y cells and is therefore considered to be non-mutagenic under the conditions of the test. - Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) and the source substance FATTY ACIDS, C16-18, ISOBUTYL ESTERS (CAS 85865-69-6) are both Short Chain Alcohol Esters (SCAE C2-C8) composed by a fatty acid (C16-C18) and a C4 alcohol (isobutanol).
The source and the target substance show therefore the same reactive groups and a similar composition. A read-across to the source is therefore justified.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Both target and source substances are fatty acid esters produced by chemical reaction of an alcohol (isobutanol) with organic acids (e. g. stearic acid) in the presence of an acid catalyst. The esterification reaction is started by a transfer of a proton from the acid catalyst to the acid to form an alkyloxonium ion. The carboxylic acid is protonated on its carbonyl oxygen followed by a nucleophilic addition of a molecule of the alcohol to a carbonyl carbon of acid. An intermediate product is formed. This intermediate product loses a water molecule and proton to give an ester. Monoesters are the final product of esterification.
3. ANALOGUE APPROACH JUSTIFICATION
Since both target and source substances are fatty acid esters produced by chemical reaction of an alcohol (isobutanol) with an organic acid and therefore share similar/overlapping structural features and functional groups, it is justified to use a read across approach. The source substance has been registered already and its genetic toxicity in vitro has been investigated using a grouping of substance and read across approach. The mutagenic potential of the source substance, Fatty acids, C16-18, Isobutyl Esters (CAS# 85865-69-6), was tested in a Salmonella typhimurium reverse mutation assay equivalent to OECD Guideline 471 and under GLP. Fatty acids, C16-18, Isobutyl Esters did not induce point mutations by base-pair changes or frame-shifts in the genome of the strains used.
All available data investigating the genetic toxicity indicate that members of the category Fatty acid C2-8 esters have no genotoxic potential and classification according to EU classification criteria for genetic toxicity is not required.
The same behaviour is predicted for the target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) . - Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- 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:
- no 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:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- 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:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Conclusions:
- Fatty acids, C16-18, Isobutyl Esters did not induce point mutations by base-pair changes or frame-shifts in the genome of the strains used.
All available data investigating the genetic toxicity indicate that members of the category Fatty acid C2-8 esters have no genotoxic potential and classification according to EU classification criteria for genetic toxicity is not required.
The same behaviour is predicted for the target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) . - Executive summary:
The target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) and the source substance FATTY ACIDS, C16-18, ISOBUTYL ESTERS (CAS 85865-69-6) are both Short Chain Alcohol Esters (SCAE C2-C8) composed by a fatty acid (C16-C18) and a C4 alcohol (isobutanol).
The source and the target substance show therefore the same reactive groups and a similar composition. A read-across to the source is therefore justified.
The source substance has been registered already and its GENETIC TOXICITY potential has been investigated using a grouping of substance and read across approach. The mutagenic potential of the source substance, Fatty acids, C16-18, Isobutyl Esters (CAS# 85865-69-6), was tested in a Salmonella typhimurium reverse mutation assay equivalent to OECD Guideline 471 and under GLP. Fatty acids, C16-18, Isobutyl Esters did not induce point mutations by base-pair changes or frame-shifts in the genome of the strains used.
All available data investigating the genetic toxicity indicate that members of the category Fatty acid C2-8 esters have no genotoxic potential and classification according to EU classification criteria for genetic toxicity is not required.
The same behaviour is predicted for the target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) .
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) and the source substance FATTY ACIDS, C16-18, ISOBUTYL ESTERS (CAS 85865-69-6) are both Short Chain Alcohol Esters (SCAE C2-C8) composed by a fatty acid (C16-C18) and a C4 alcohol (isobutanol).
The source and the target substance show therefore the same reactive groups and a similar composition. A read-across to the source is therefore justified.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Both target and source substances are fatty acid esters produced by chemical reaction of an alcohol (isobutanol) with organic acids (e. g. stearic acid) in the presence of an acid catalyst. The esterification reaction is started by a transfer of a proton from the acid catalyst to the acid to form an alkyloxonium ion. The carboxylic acid is protonated on its carbonyl oxygen followed by a nucleophilic addition of a molecule of the alcohol to a carbonyl carbon of acid. An intermediate product is formed. This intermediate product loses a water molecule and proton to give an ester. Monoesters are the final product of esterification.
3. ANALOGUE APPROACH JUSTIFICATION
Since both target and source substances are fatty acid esters produced by chemical reaction of an alcohol (isobutanol) with an organic acid and therefore share similar/overlapping structural features and functional groups, it is justified to use a read across approach. The source substance has been registered already and its cytogenicity in vitro has been investigated using a grouping of substance and read across approach. The clastogenic potential of the source substance, Fatty acids, C16-18, Isobutyl Esters (CAS# 85865-69-6), was tested in an in vitro mammalian chromosome aberration test. Fatty acids, C16-18, Isobutyl Esters did not show clastogenic effects.
All available data investigating the genetic toxicity indicate that members of the category Fatty acid C2-8 esters have no clastogenic potential and classification according to EU classification criteria for genetic toxicity is not required.
The same behaviour is predicted for the target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) . - Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- primary culture, other: human lymphocytes
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- max 38% inhibition at 468,75 mg/mL
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- primary culture, other: human lymphocytes
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Remarks:
- max 11% (1st experiment) and 22% (2nd experiment)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Negative non-clastogenic to human lymphocytes in vitro
The numbers of chromosome aberrations and polyploid cells were not increased at any dose level in comparison to the control. The test substance is non-clastogenic in mammalian cells in vitro. - Executive summary:
The target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) and the source substance FATTY ACIDS, C16-18, ISOBUTYL ESTERS (CAS 85865-69-6) are both Short Chain Alcohol Esters (SCAE C2-C8) composed by a fatty acid (C16-C18) and a C4 alcohol (isobutanol).
The source substance has been registered already and its cytogenicity in vitro has been investigated using a grouping of substance and read across approach. The clastogenic potential of the source substance, Fatty acids, C16-18, Isobutyl Esters (CAS# 85865-69-6), was tested in an in vitro mammalian chromosome aberration test. Fatty acids, C16-18, Isobutyl Esters did not show clastogenic effects.
All available data investigating the genetic toxicity indicate that members of the category Fatty acid C2-8 esters have no clastogenic potential and classification according to EU classification criteria for genetic toxicity is not required.
The same behaviour is predicted for the target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) .
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) and the source substance FATTY ACIDS, C16-18, ISOBUTYL ESTERS (CAS 85865-69-6) are both Short Chain Alcohol Esters (SCAE C2-C8) composed by a fatty acid (C16-C18) and a C4 alcohol (isobutanol).
The source and the target substance show therefore the same reactive groups and a similar composition. A read-across to the source is therefore justified.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Both target and source substances are fatty acid esters produced by chemical reaction of an alcohol (isobutanol) with organic acids (e. g. stearic acid) in the presence of an acid catalyst. The esterification reaction is started by a transfer of a proton from the acid catalyst to the acid to form an alkyloxonium ion. The carboxylic acid is protonated on its carbonyl oxygen followed by a nucleophilic addition of a molecule of the alcohol to a carbonyl carbon of acid. An intermediate product is formed. This intermediate product loses a water molecule and proton to give an ester. Monoesters are the final product of esterification.
3. ANALOGUE APPROACH JUSTIFICATION
Since both target and source substances are fatty acid esters produced by chemical reaction of an alcohol (isobutanol) with an organic acid and therefore share similar/overlapping structural features and functional groups, it is justified to use a read across approach. The source substance has been registered already and its cytogenicity in vitro has been investigated using a grouping of substance and read across approach. The clastogenic potential of the source substance, Fatty acids, C16-18, Isobutyl Esters (CAS# 85865-69-6), was tested in an in vitro mammalian chromosome aberration test. Fatty acids, C16-18, Isobutyl Esters did not show clastogenic effects.
All available data investigating the genetic toxicity indicate that members of the category Fatty acid C2-8 esters have no clastogenic potential and classification according to EU classification criteria for genetic toxicity is not required.
The same behaviour is predicted for the target substance FATTY ACIDS, COCO, ISO-BU ESTERS (CAS 91697-43-7) . - Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
Referenceopen allclose all
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) |
|||||||||
Base-pair substitution type |
Frameshift type |
||||||||||
TA 100 |
TA1535 |
TA1538 |
TA98 |
TA1537 |
|||||||
– |
Buffer control |
77.3 |
7.0 |
9.7 |
29.0 |
5.3 |
|||||
– |
Solvent control |
94.7 |
7.7 |
8.3 |
27.3 |
5.3 |
|||||
– |
8 |
82.0 |
7.7 |
9.0 |
29.0 |
5.0 |
|||||
– |
40 |
92.3 |
10.3 |
13.0 |
32.3 |
4.3 |
|||||
– |
200 |
84.0 |
7.3 |
10.0 |
27.0 |
8.0 |
|||||
– |
1000 |
88.0 |
6.7 |
8.0 |
32.7 |
4.3 |
|||||
– |
5000 |
80.7 |
9.0 |
11.7 |
30.7 |
5.3 |
|||||
Positive controls, –S9 |
Name |
SA |
2-AA |
SA |
2-AA |
4-NoP |
2-AA |
4-NoP |
2-AA |
9-AA |
2-AA |
Concentrations (μg/plate) |
2 |
5 |
2 |
2.5 |
40 |
5 |
40 |
5 |
80 |
2.5 |
|
Mean No. of colonies/plate (average of 3) |
314 |
79.3 |
393 |
7.0 |
1488 |
15 |
587 |
35 |
537 |
3.7 |
|
+ |
Buffer control |
87.7 |
9.7 |
12.0 |
25.0 |
9.3 |
|||||
+ |
Solvent control |
90.3 |
11.3 |
15.0 |
31.3 |
7.3 |
|||||
+ |
8 |
85.3 |
7.0 |
14.7 |
36.7 |
6.0 |
|||||
+ |
40 |
92.3 |
11.0 |
12.3 |
31.3 |
6.3 |
|||||
+ |
200 |
88.0 |
9.7 |
15.0 |
38.7 |
4.7 |
|||||
+ |
1000 |
94.7 |
12.0 |
12.0 |
33.0 |
6.0 |
|||||
+ |
5000 |
99.7 |
11.7 |
14.0 |
35.0 |
6.3 |
|||||
Positive controls, +S9 |
Name |
SA |
2-AA |
SA |
2-AA |
4-NoP |
2-AA |
4-NoP |
2-AA |
9-AA |
2-AA |
Concentrations (μg/plate) |
2 |
5 |
2 |
2.5 |
40 |
5 |
40 |
5 |
80 |
2.5 |
|
Mean No. of colonies/plate (average of 3) |
223 |
1212 |
122 |
150 |
1113 |
1475 |
576 |
1104 |
281 |
127 |
|
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) |
|||||||||
Base-pair substitution type |
Frameshift type |
||||||||||
TA 100 |
TA1535 |
TA1538 |
TA98 |
TA1537 |
|||||||
– |
Buffer control |
82.0 |
7.7 |
11.7 |
30.7 |
4.3 |
|||||
– |
Solvent control |
91.0 |
9.3 |
15.0 |
30.3 |
4.3 |
|||||
– |
8 |
80.7 |
8.3 |
14.7 |
27.0 |
7.0 |
|||||
– |
40 |
86.0 |
8.3 |
16.3 |
30.0 |
6.3 |
|||||
– |
200 |
77.0 |
6.7 |
15.7 |
34.7 |
4.0 |
|||||
– |
1000 |
89.7 |
7.3 |
16.3 |
35.3 |
6.0 |
|||||
– |
5000 |
91.3 |
8.7 |
11.3 |
33.7 |
4.7 |
|||||
Positive controls, –S9 |
Name |
SA |
2-AA |
SA |
2-AA |
4-NoP |
2-AA |
4-NoP |
2-AA |
9-AA |
2-AA |
Concentrations (μg/plate) |
2 |
5 |
2 |
2.5 |
40 |
5 |
40 |
5 |
80 |
2.5 |
|
Mean No. of colonies/plate (average of 3) |
295 |
77 |
288 |
5 |
1516 |
20 |
681 |
21 |
274 |
5 |
|
+ |
Buffer control |
93.0 |
9.0 |
18.0 |
28.7 |
7.0 |
|||||
+ |
Solvent control |
96.0 |
8.0 |
13.7 |
24.3 |
8.3 |
|||||
+ |
8 |
85.3 |
8.3 |
19.7 |
29.0 |
5.7 |
|||||
+ |
40 |
93.3 |
6.7 |
17.7 |
36.7 |
6.3 |
|||||
+ |
200 |
87.7 |
11.3 |
16.0 |
25.0 |
6.7 |
|||||
+ |
1000 |
106.3 |
9.0 |
17.0 |
30.7 |
5.7 |
|||||
+ |
5000 |
100.3 |
10.7 |
15.3 |
31.3 |
9.0 |
|||||
Positive controls, +S9 |
Name |
SA |
2-AA |
SA |
2-AA |
4-NoP |
2-AA |
4-NoP |
2-AA |
9-AA |
2-AA |
Concentrations (μg/plate) |
2 |
5 |
2 |
2.5 |
40 |
5 |
40 |
5 |
80 |
2.5 |
|
Mean No. of colonies/plate (average of 3) |
233 |
1176 |
98 |
123 |
1268 |
1228 |
786 |
1092 |
306 |
126 |
|
SA = sodium azide
2-AA = 2 -aminoanthracene
4 -NoP = 4 -nitro-o-phenylenediamine
9 -AA = 9-aminoacridine
Table 1: Mean values of chromosome aberrations and polyploid cells
|
Experiment 1 |
Experiment 2 |
||||||
|
24h without S9 |
4h with S9 |
4h without S9 |
4h with S9 |
||||
Dose level (µg/mL) |
% cells with aberrations (-gaps) |
% cells with polyploids |
% cells with aberrations (-gaps) |
% cells with polyploids |
% cells with aberrations (-gaps) |
% cells with polyploids |
% cells with aberrations (-gaps) |
% cells with polyploids |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0.5 |
0.5 |
312.5 |
0.5 |
0 |
- |
- |
- |
- |
- |
- |
468.75 |
0.5 |
0 |
- |
- |
- |
- |
- |
- |
625 |
1.5 |
0 |
1.0 |
0 |
0 |
0 |
0.5 |
0 |
1250 |
- |
- |
1.0 |
0.5 |
0.5 |
0 |
0 |
0 |
2500 |
- |
- |
0 |
0 |
0 |
0 |
0 |
0 |
positive control |
54.0 |
0 |
39.3 |
0 |
42.0 |
0 |
28.7 |
0 |
Table 1: Results for the Relative Suspension Growth (%RSG) in the preliminary toxicity test were as follows:
Dose (µg/mL) |
% RSG (-S9) 4-Hour Exposure |
% RSG (+S9) 4-Hour Exposure |
% RSG (-S9) 24-Hour Exposure |
0 |
100 |
100 |
100 |
19.53 |
88 |
84 |
61 |
39.06 |
83 |
92 |
61 |
78.13 |
74 |
92 |
56 |
156.25 |
91 |
78 |
58 |
312.5 |
85 |
78 |
53 |
625 |
75 |
64 |
55 |
1250 |
84 |
84 |
50 |
2500 |
85 |
77 |
44 |
5000 |
65 |
91 |
64 |
Table 2: Results from Experiment 1
Treatment (µg/mL) |
4-Hours without S9 |
Treatment (µg/mL) |
4-Hours with S9 |
||||
%RSG |
RTG |
MF§ |
%RSG |
RTG |
MF§ |
||
0 |
100 |
1.00 |
100.35 |
0 |
100 |
1.00 |
122.76 |
156.25 |
130 |
1.34 |
96.34 |
156.25 |
98 |
1.05 |
88.56 |
312.5 |
112 |
1.28 |
98.69 |
312.5 |
92 |
1.00 |
106.06 |
625 |
136 |
1.57 |
91.80 |
625 |
82 |
0.86 |
137.37 |
1250 |
129 |
1.20 |
138.71 |
1250 |
94 |
0.92 |
122.97 |
2500 |
127 |
1.47 |
82.69 |
2500 |
102 |
0.87 |
110.73 |
5000 |
152 |
1.43 |
119.09 |
5000 |
109 |
0.95 |
112.66 |
Linear Trend NS |
Linear Trend NS |
||||||
EMS 400 |
124 |
0.83 |
748.75 |
CP 2 |
94 |
0.51 |
951.61 |
Table 3: Results from Experiment 2
Treatment (µg/mL) |
24-Hours without S9 |
Treatment (µg/mL) |
4-Hours with S9 |
||||
%RSG |
RTG |
MF§ |
%RSG |
RTG |
MF§ |
||
0 |
100 |
1.00 |
88.69 |
0 |
100 |
1.00 |
131.27 |
156.25 |
63 |
0.69 |
95.98 |
156.25 |
47 |
0.41 |
210.65 |
312.5 |
66 |
0.62 |
57.47 |
312.5 |
58 |
0.46 |
229.00 |
625 |
72 |
0.77 |
99.53 |
625 |
35 |
0.33 |
213.23 |
1250 |
63 |
0.61 |
64.44 |
1250 |
88 |
0.82 |
150.73 |
2500 |
76 |
0.81 |
82.48 |
2500 |
113 |
1.03 |
128.99 |
5000 |
99 |
1.l2 |
90.36 |
5000 |
127 |
1.09 |
133.50 |
Linear Trend NS |
Linear Trend NS |
||||||
EMS 400 |
103 |
0.73 |
1021.46 |
CP 2 |
67 |
0.527 |
1305.55 |
Table 4: Results from Experiment 3
Treatment (µg/mL) |
4-Hours withS9 |
||
%RSG |
RTG |
MF§ |
|
0 |
100 |
1.00 |
165.58 |
39.06 |
110 |
0.98 |
151.66 |
78.13 |
101 |
1.03 |
185.56 |
117.2 |
61 |
0.41 |
167.55 |
156.25 |
50 |
0.39 |
170.51 |
234.38 |
138 |
1.32 |
174.72 |
312.5 |
124 |
1.32 |
128.11 |
468.75 |
64 |
0.63 |
176.42 |
625 |
60 |
0.41 |
228.91 |
937.5 |
63 |
0.46 |
190.85 |
1250 |
65 |
0.46 |
156.75 |
Linear Trend NS |
|||
CP 2 |
62 |
0.22 |
856.01 |
%RSG: Relative Suspension Growth
RTG : Relative Total Growth
MF§ : 5-TFT resistant mutants/10^6 viable cells 2 days after treatment
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: gene mutation
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- other:
Reference
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Justification for classification or non-classification
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