Registration Dossier
Registration Dossier
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 285-203-4 | CAS number: 85049-34-9
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
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 05 Jun - 09 Aug 1991
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Remarks:
- GLP - Guideline study with acceptable restrictions. No S. typhimurium TA102 or E. coli strain was included.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- adopted May 26, 1983 (2)
- Deviations:
- yes
- Remarks:
- no S. typhimurium TA102 or E. coli strain was included
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- adopted March 2, 1983
- Deviations:
- yes
- Remarks:
- no S. typhimurium TA102 or E. coli strain was included
- 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
- Species / strain / cell type:
- S. typhimurium TA 1538
- 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:
- First and second experiment: 8, 40, 200, 1000 and 5000 µg/plate with and without metabolic activation
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Suspension medium Tween 80/bidest. water
- Justification for choice of solvent/vehicle: According to the author, the suspension medium was chosen based on the solubility properties preliminary tested before start of the study. - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Tween 80/bidest. water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: sodium azide (2 µg/plate, ±S9, TA1535 and TA100); 9-Aminoacridine (80 µg/plate,±S9, TA1537); 4-Nitro-o-phenylendiamine (40 µg/plate, ±S9,TA98 and TA1538); 2 Amino-anthracene (2.5 or 5 µg/plate, ±S9, TA1535 and TA1357; TA100, TA1538 and TA98)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h
NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments - Evaluation criteria:
- The test material may be considered positive in this test system if a combination of the following criteria are met:
- the plate background of non-reverted bacteria does not show any growth reduction versus the respective negative controls.
- the spontaneous mutation rates of each tester strain per plate are within the characteristic spontaneous mutation range.
- the positive controls show mutation rates exceeding the control values of TA100 at least two fold and those of the other strains at least by the factor 3.
- at more than one dose tested, at least a two-fold (or more) increase in comparison with the negative controls in the tester strain TA100. For the other strains, an increase in the mutation rate of more than 3 above the corresponding negative controls. - Statistics:
- Mean values and standard deviation were calculated.
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- 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 nor precipitates, but tested up to recommended limit concentrations
- 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 nor precipitates, but tested up to recommended limit concentrations
- 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 nor precipitates, but tested up to recommended limit concentrations
- 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 nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Other confounding effects: Contamination of 1 plate of strain TA98 in the second experiment (1000 µg/plate), therefore only 2 individual values were used for calculation of the mean number of revertant colonies at this concentration.
COMPARISON WITH HISTORICAL CONTROL DATA:
- see Table 1 and Table 2 under "Any other information on results incl. tables". - Remarks on result:
- other: all strains/cell types tested
- Conclusions:
- Interpretation of results: negative
- 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
- Study period:
- 05 Jun - 09 Aug 1991
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Remarks:
- GLP - Guideline study with acceptable restrictions. No S. typhimurium TA102 or E. coli strain was included.
- Justification for type of information:
- Please refer section 13 for read across justification.
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- adopted May 26, 1983 (2)
- Deviations:
- yes
- Remarks:
- no S. typhimurium TA102 or E. coli strain was included
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- adopted March 2, 1983
- Deviations:
- yes
- Remarks:
- no S. typhimurium TA102 or E. coli strain was included
- 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
- Species / strain / cell type:
- S. typhimurium TA 1538
- 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:
- First and second experiment: 8, 40, 200, 1000 and 5000 µg/plate with and without metabolic activation
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Suspension medium Tween 80/bidest. water
- Justification for choice of solvent/vehicle: According to the author, the suspension medium was chosen based on the solubility properties preliminary tested before start of the study. - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Tween 80/bidest. water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: sodium azide (2 µg/plate, ±S9, TA1535 and TA100); 9-Aminoacridine (80 µg/plate,±S9, TA1537); 4-Nitro-o-phenylendiamine (40 µg/plate, ±S9,TA98 and TA1538); 2 Amino-anthracene (2.5 or 5 µg/plate, ±S9, TA1535 and TA1357; TA100, TA1538 and TA98)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h
NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments - Evaluation criteria:
- The test material may be considered positive in this test system if a combination of the following criteria are met:
- the plate background of non-reverted bacteria does not show any growth reduction versus the respective negative controls.
- the spontaneous mutation rates of each tester strain per plate are within the characteristic spontaneous mutation range.
- the positive controls show mutation rates exceeding the control values of TA100 at least two fold and those of the other strains at least by the factor 3.
- at more than one dose tested, at least a two-fold (or more) increase in comparison with the negative controls in the tester strain TA100. For the other strains, an increase in the mutation rate of more than 3 above the corresponding negative controls. - Statistics:
- Mean values and standard deviation were calculated.
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- 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 nor precipitates, but tested up to recommended limit concentrations
- 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 nor precipitates, but tested up to recommended limit concentrations
- 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 nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Other confounding effects: Contamination of 1 plate of strain TA98 in the second experiment (1000 µg/plate), therefore only 2 individual values were used for calculation of the mean number of revertant colonies at this concentration.
COMPARISON WITH HISTORICAL CONTROL DATA:
- see Table 1 and Table 2 under "Any other information on results incl. tables". - Remarks on result:
- other: all strains/cell types tested
- Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1997
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Remarks:
- Comparable to guideline study with acceptable restrictions. No discussion of results and no historical control data.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- no discussion of results and no historical control data.
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- no discussion of results and no historical control data.
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Ministerium für Umwelt, Raumordnung und Landwirtschaft des Landes Nordrhein-Westfalen, Düsseldorf, Germany
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Not applicable
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media:
MEM medium supplemented with 2 mM L-Glutamine and
- 4% (v/v) fetal calf serum (MEM4) or
- 0% (v/v) fetal calf serum (MEM0)
- 100 IU/mL penicillin/streptomycin
During exposure to the test substancewith S9 mix, MEM0 medium was used and replaced by MEM4 after 3 h after test substance administration. - Additional strain / cell type characteristics:
- other: modal chromosome number of 22 and a cell cycle length of approx. 16.5 h
- 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:
- 18h treatment: 10, 40 and 80 µg/mL (without metabolic activation)
18h treatment: 10, 60, 80 and 100 µg/mL (with metabolic activation)
28h treatment: 80 µg/mL (without metabolic activation)
28h treatment: 100 µg/mL (with metabolic activation) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- MEM4 medium (resp. MEM0 medium in the test with S9 mix) containing 1% (v/v) ethanol
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: cyclophosphamide, 3 and 4 µg/mL in MEM0 medium, +S9; mitomycin C, 0.03 and 0.04 µg/mL in MEM4 medium, -S9
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 18 and 28 h
- Fixation time (start of exposure up to fixation or harvest of cells): 18 h treatment: 18 h; 28 h treatment: 28 h
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: 2 replications each in one (28 h treatment) or two (18 h treatment) independent experiment
NUMBER OF CELLS EVALUATED: 100 per culture
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index of 1000 cells per slide
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreduplicated cells: yes - Evaluation criteria:
- The test chemical is to be considered clastogenic in this assay if:
- it induces chromosomal aberrations (excl. gaps) in a statistically significant manner in one or more concentrations
- the induced proportion of aberrant cells at such test substance concentrations exceeds the normal range of the test system
- positive results can be verified in an independent experiment. - Statistics:
- The number of aberrant cells in each replica was used to establish acceptable homogeneity between replicates by means of a binomial dispersion test ( Richardson, C., Williams, D. A., Allen, J. A., Amphlett, G., Chanter, D. O. and Phillips, B. Analysis of Data from In Vitro Cytogenetic Assays, in: Statistical Evaluation of Mutagenicity Test Data, Kirkland, D. J., (ed) Cambridge University Press, Cambridge, pp. 41-64., 1990 ). The proportion of cells that was treated with the test substance and harboured structural aberrations (excl. gaps) was compared with the corresponding proportion of the negative controls in the Chi-square test. Probability values of p < 0.05 were accepted as statistically significant.
- Key result
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- systematic influence of the test compound which led to a reduction in the mitotic index from 10 µg/mL.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: The test substance was soluble in ethanol, in MEM4 medium containing 1% ethanol the solubility limit of the test substance was determined to be 100 µg/mL (homogeneous emulsion).
- Other confounding effects: In the first test without metabolic activation the negative controls exhibited only a mitotic index of 2.0%. Therefore, test 1 without metabolic activation was completely repeated with the same doses and named test #1a. - Remarks on result:
- other: all strains/cell types tested
- Conclusions:
- Interpretation of results: negative
- 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
- Study period:
- 1997
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Remarks:
- Comparable to guideline study with acceptable restrictions. No discussion of results and no historical control data.
- Justification for type of information:
- Please refer Section 13 for read across justification.
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- no discussion of results and no historical control data.
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- no discussion of results and no historical control data.
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Ministerium für Umwelt, Raumordnung und Landwirtschaft des Landes Nordrhein-Westfalen, Düsseldorf, Germany
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Not applicable
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media:
MEM medium supplemented with 2 mM L-Glutamine and
- 4% (v/v) fetal calf serum (MEM4) or
- 0% (v/v) fetal calf serum (MEM0)
- 100 IU/mL penicillin/streptomycin
During exposure to the test substancewith S9 mix, MEM0 medium was used and replaced by MEM4 after 3 h after test substance administration. - Additional strain / cell type characteristics:
- other: modal chromosome number of 22 and a cell cycle length of approx. 16.5 h
- 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:
- 18h treatment: 10, 40 and 80 µg/mL (without metabolic activation)
18h treatment: 10, 60, 80 and 100 µg/mL (with metabolic activation)
28h treatment: 80 µg/mL (without metabolic activation)
28h treatment: 100 µg/mL (with metabolic activation) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- MEM4 medium (resp. MEM0 medium in the test with S9 mix) containing 1% (v/v) ethanol
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: cyclophosphamide, 3 and 4 µg/mL in MEM0 medium, +S9; mitomycin C, 0.03 and 0.04 µg/mL in MEM4 medium, -S9
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 18 and 28 h
- Fixation time (start of exposure up to fixation or harvest of cells): 18 h treatment: 18 h; 28 h treatment: 28 h
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: 2 replications each in one (28 h treatment) or two (18 h treatment) independent experiment
NUMBER OF CELLS EVALUATED: 100 per culture
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index of 1000 cells per slide
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreduplicated cells: yes - Evaluation criteria:
- The test chemical is to be considered clastogenic in this assay if:
- it induces chromosomal aberrations (excl. gaps) in a statistically significant manner in one or more concentrations
- the induced proportion of aberrant cells at such test substance concentrations exceeds the normal range of the test system
- positive results can be verified in an independent experiment. - Statistics:
- The number of aberrant cells in each replica was used to establish acceptable homogeneity between replicates by means of a binomial dispersion test ( Richardson, C., Williams, D. A., Allen, J. A., Amphlett, G., Chanter, D. O. and Phillips, B. Analysis of Data from In Vitro Cytogenetic Assays, in: Statistical Evaluation of Mutagenicity Test Data, Kirkland, D. J., (ed) Cambridge University Press, Cambridge, pp. 41-64., 1990 ). The proportion of cells that was treated with the test substance and harboured structural aberrations (excl. gaps) was compared with the corresponding proportion of the negative controls in the Chi-square test. Probability values of p < 0.05 were accepted as statistically significant.
- Key result
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- systematic influence of the test compound which led to a reduction in the mitotic index from 10 µg/mL.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: The test substance was soluble in ethanol, in MEM4 medium containing 1% ethanol the solubility limit of the test substance was determined to be 100 µg/mL (homogeneous emulsion).
- Other confounding effects: In the first test without metabolic activation the negative controls exhibited only a mitotic index of 2.0%. Therefore, test 1 without metabolic activation was completely repeated with the same doses and named test #1a. - Remarks on result:
- other: all strains/cell types tested
- Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 17 Jun 2010 - 30 Aug 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Remarks:
- GLP-Guideline study, tested with the source substance Fatty acids, C16-18, esters with ethylene glycol(CAS 91031-31-1). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (Dec 2012)”, 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
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- TK locus
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 1640 Hepes buffered medium (Dutch modification) containing penicillin/streptomycin, sodium pyruvate and L-glutamin amd 10% (v/v) heat-inactivated horse serum (24 h exposure); for 3 h exposure only 5% (v/v) heat-inactivated horse serum were included. Selective medium consisted of the basic medium and 20% (v/v) heat-inactivated horse serum and 5 µg/mL trifluorothymidine (TFT).
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital/β-naphtoflavone-induced rat liver S9
- Test concentrations with justification for top dose:
- Experiment 1:
With and without S9-mix: 0.1, 0.3, 1.0, 3.0, 10.0, 33.0, 100.0 and 333.0 µg/mL (3 h)
Experiment 2:
In the absence of S9-mix: 3.0, 10.0, 33.0, 100.0, 125.0, 140.0 and 175.0 µg/mL for 24h
In the presence of 12% (v/v) S9-mix: 0, 0.1, 0.3, 1.0, 3.0, 10.0, 33.0, 100.0 and 333.0 µg/mL for 3h - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: cyclophosphamide, 7.5 µg/mL, with S9; methylmethanesulfonate, 15 µg/mL and 5 µg/mL without S9, for 3 h and 24 h treatment, respectively
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 3 and 24 h
- Expression time (cells in growth medium): 48 h
SELECTION AGENT (mutation assays): 5 µg/mL trifluorothymidine
- Selection time (if incubation with a selection agent): 11-12 days
NUMBER OF REPLICATIONS: 2 replications each in 5 independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency, relative total growth, relative survival, suspension growth, relative suspension growth, growth rate - Evaluation criteria:
- A mutation assay was considered acceptable if it met the following criteria:
a) The absolute cloning efficiency of the solvent controls is between 65 and 120%. An acceptable number of surviving cells (10E6) could be analysed for expression of the TK mutation.
b) The spontaneous mutation frequency in the solvent control is ≥ 50E-06 and ≤ 170E-06.
c) The growth rate (R) over the 2-day expression period for the negative controls should be between 8 and 32 (3 h treatment) and between 32 and 180 (24 h treatment).
d) The mutation frequency of MMS should not be below 500E-06, and fro CP not below 700E-06.
The global evaluation factor (GEF) has been identified by the IWTG as the mean of the negative/solvent mutation frequeny (MF) distribution plus one standard deviation. For the micro well version of the assay the GEF is 126.
A test substance is considered positive (mutagenic) in the mutation assay if it induces a MF of more than the MF of the controls + 126 in a dose-dependent manner. An observed increase should be biologically relevant and will be compared with the historical control data range.
A test substance is considered equivocal (questionable) in the mutation assay if no clear conclusion for positive or negative result can be made after an additional confirmation study.
A test substance is considered negative (not mutagenic) in the mutation assay if: a) None of the tested concentrations reaches a mutation frequency of the MF of the controls + 126. b) The results are confirmed in an independently repeated test. - Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility and precipitation: The test substance precipitated in the exposure medium at concentrations of 100 µg/mL and above. The test substance was tested beyond the limit of the solubility to obtain adequate cytotoxicity data, the concentration used as the highest test substance concentration for the dose range ending test was 333 µg/mL.
RANGE-FINDING/SCREENING STUDIES:
In the absence of S9-mix, the relative suspension growth was 66% at the test substance concentration of 333 µg/mL compared to the relative suspension growth of the solvent control. In the presence of S9-mix, no toxicity in the relative suspension growth was observed up to and including the highest test substance concentration of 333 µg/mL compared to the suspension growth of the solvent control.
COMPARISON WITH HISTORICAL CONTROL DATA: The spontaneous mutation frequencies in the solvent treated control cultures were within the ranges of the historical controls. - Remarks on result:
- other: strain/cell type:
- Conclusions:
- Interpretation of results: negative
- 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
- Study period:
- 17 Jun 2010 - 30 Aug 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Remarks:
- GLP-Guideline study, tested with the source substance Fatty acids, C16-18, esters with ethylene glycol(CAS 91031-31-1). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (Dec 2012)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
- Justification for type of information:
- Please refer section 13 for read across justification.
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- TK locus
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 1640 Hepes buffered medium (Dutch modification) containing penicillin/streptomycin, sodium pyruvate and L-glutamin amd 10% (v/v) heat-inactivated horse serum (24 h exposure); for 3 h exposure only 5% (v/v) heat-inactivated horse serum were included. Selective medium consisted of the basic medium and 20% (v/v) heat-inactivated horse serum and 5 µg/mL trifluorothymidine (TFT).
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital/β-naphtoflavone-induced rat liver S9
- Test concentrations with justification for top dose:
- Experiment 1:
With and without S9-mix: 0.1, 0.3, 1.0, 3.0, 10.0, 33.0, 100.0 and 333.0 µg/mL (3 h)
Experiment 2:
In the absence of S9-mix: 3.0, 10.0, 33.0, 100.0, 125.0, 140.0 and 175.0 µg/mL for 24h
In the presence of 12% (v/v) S9-mix: 0, 0.1, 0.3, 1.0, 3.0, 10.0, 33.0, 100.0 and 333.0 µg/mL for 3h - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: cyclophosphamide, 7.5 µg/mL, with S9; methylmethanesulfonate, 15 µg/mL and 5 µg/mL without S9, for 3 h and 24 h treatment, respectively
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 3 and 24 h
- Expression time (cells in growth medium): 48 h
SELECTION AGENT (mutation assays): 5 µg/mL trifluorothymidine
- Selection time (if incubation with a selection agent): 11-12 days
NUMBER OF REPLICATIONS: 2 replications each in 5 independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency, relative total growth, relative survival, suspension growth, relative suspension growth, growth rate - Evaluation criteria:
- A mutation assay was considered acceptable if it met the following criteria:
a) The absolute cloning efficiency of the solvent controls is between 65 and 120%. An acceptable number of surviving cells (10E6) could be analysed for expression of the TK mutation.
b) The spontaneous mutation frequency in the solvent control is ≥ 50E-06 and ≤ 170E-06.
c) The growth rate (R) over the 2-day expression period for the negative controls should be between 8 and 32 (3 h treatment) and between 32 and 180 (24 h treatment).
d) The mutation frequency of MMS should not be below 500E-06, and fro CP not below 700E-06.
The global evaluation factor (GEF) has been identified by the IWTG as the mean of the negative/solvent mutation frequeny (MF) distribution plus one standard deviation. For the micro well version of the assay the GEF is 126.
A test substance is considered positive (mutagenic) in the mutation assay if it induces a MF of more than the MF of the controls + 126 in a dose-dependent manner. An observed increase should be biologically relevant and will be compared with the historical control data range.
A test substance is considered equivocal (questionable) in the mutation assay if no clear conclusion for positive or negative result can be made after an additional confirmation study.
A test substance is considered negative (not mutagenic) in the mutation assay if: a) None of the tested concentrations reaches a mutation frequency of the MF of the controls + 126. b) The results are confirmed in an independently repeated test. - Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility and precipitation: The test substance precipitated in the exposure medium at concentrations of 100 µg/mL and above. The test substance was tested beyond the limit of the solubility to obtain adequate cytotoxicity data, the concentration used as the highest test substance concentration for the dose range ending test was 333 µg/mL.
RANGE-FINDING/SCREENING STUDIES:
In the absence of S9-mix, the relative suspension growth was 66% at the test substance concentration of 333 µg/mL compared to the relative suspension growth of the solvent control. In the presence of S9-mix, no toxicity in the relative suspension growth was observed up to and including the highest test substance concentration of 333 µg/mL compared to the suspension growth of the solvent control.
COMPARISON WITH HISTORICAL CONTROL DATA: The spontaneous mutation frequencies in the solvent treated control cultures were within the ranges of the historical controls. - Remarks on result:
- other: strain/cell type:
- Conclusions:
- Interpretation of results: negative
Referenceopen allclose all
COMPARISON WITH HISTORICAL DATA
Table 1. Characteristic spontaneous mutation range of the test batches without S9-mix.
Tester strains |
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
Historical laboratory values (Mean values) Extreme values |
6 1-25 |
87 35-201 |
7 1-24 |
15 6-27 |
20 5-39 |
Ames et al.* (Mean values) Extreme values |
20 5-50 |
160 60-220 |
7 3-25 |
25 5-40 |
40 15-75 |
*Ames, BN, McCann, J, Yamasaki, E 1977, ´Methods for detecting carcinogens and mutagens with the Salmonella/mammalian microsome mutagenicity test.`in Kilbey et al. Handbook of Mutagenicity Test Procedures, Elsevier, Amsterdam, pp. 1 -17.
Table 2. Characteristic spontaneous mutation range of the test batches containing S9-mix.
Tester strains |
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
Historical laboratory values (Mean values) Extreme values |
8 1-25 |
105 54-252 |
6 1-23 |
18 3-48 |
27 7-76 |
STUDY RESULTS
Table 3. 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 |
|||||
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
||
– |
Negative control |
9.7 ± 3.1 |
97.7 ± 2.1 |
6.3 ± 3.2 |
6.7 ± 1.2 |
46.7 ± 3.2 |
– |
Solvent control |
7.7 ± 3.5 |
94.3 ± 17.4 |
8.0 ± 1.7 |
7.3 ± 2.3 |
46.3 ± 6.0 |
– |
8 |
8.3 ± 2.1 |
70.0 ± 7.8 |
9.0 ± 1.0 |
6.7 ± 1.2 |
33.7 ± 0.6 |
– |
40 |
11.0 ± 2.7 |
74.3 ± 3.2 |
7.3 ± 2.1 |
5.3 ± 0.6 |
40.3 ± 4.9 |
– |
200 |
12.0 ± 4.6 |
67.3 ± 11.2 |
6.3 ± 1.2 |
5.7 ± 1.2 |
37.3 ± 3.2 |
– |
1000 |
10.0 ± 4.0 |
62.7 ± 10.8 |
10.3 ± 1.2 |
4.3 ± 1.5 |
42.7 ± 3.1 |
– |
5000 |
9.0 ± 1.7 |
83.0 ± 6.9 |
5.7 ± 1.5 |
4.3 ± 0.6 |
48.3 ± 3.5 |
Positive controls, –S9 |
Name |
SA |
SA |
9AA |
4NPD |
4NPD |
Concentrations (μg/plate) |
2 |
2 |
80 |
40 |
40 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
744.7 ± 49.3 |
894.0 ± 62.0 |
939.7 ± 181.4 |
1541.0 ± 96.0 |
1410.7 ± 154.4 |
|
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
|
Concentrations (μg/plate) |
2.5 |
5 |
2.5 |
5 |
5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
9.0 ± 1.7 |
88.7 ± 17.7 |
7.0 ± 2.7 |
10.3 ± 1.5 |
46.0 ± 8.7 |
|
+ |
Negative control |
5.7 ± 0.6 |
80.3 ± 15.3 |
8.7 ± 2.9 |
11.3 ± 2.3 |
53.3 ± 3.2 |
+ |
Solvent control |
9.7 ± 4.6 |
80.3 ± 14.4 |
9.3 ± 1.2 |
13.7 ± 2.3 |
44.3 ± 6.7 |
+ |
8 |
12.3 ± 3.1 |
82.3 ± 12.1 |
7.7 ± 3.5 |
14.7 ± 2.9 |
49.7 ± 4.2 |
+ |
40 |
11.3 ± 4.6 |
92.0 ± 11.8 |
6.3 ± 0.6 |
13.0 ± 4.2 |
47.7 ± 4.0 |
+ |
200 |
8.0 ± 2.0 |
84.3 ± 4.9 |
8.0 ± 1.0 |
16.3 ± 1.5 |
53.3 ± 4.0 |
+ |
1000 |
10.0 ± 2.0 |
70.7 ± 9.7 |
9.3 ± 3.8 |
13.0 ± 1.0 |
50.5 ± 3.5C |
+ |
5000 |
10.0 ± 3.6 |
74.7 ± 11.0 |
7.0 ± 1.7 |
13.0 ± 3.0 |
54.0 ± 7.0 |
Positive controls, +S9 |
Name |
SA |
SA |
9AA |
4NPD |
4NPD |
Concentrations (μg/plate) |
2 |
2 |
80 |
40 |
40 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
213.3 ± 19.8 |
230.0 ± 4.6 |
283.3 ± 27.8 |
1434.0 ± 68.9 |
1307.6 ± 68.3 |
|
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
|
Concentrations (μg/plate) |
2.5 |
5 |
2.5 |
5 |
5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
186.3 ± 19.4 |
1328.3 ± 166.5 |
52.3 ± 1.5 |
1711.3 ± 25.0 |
1587.7 ± 237.7 |
|
SA = sodium azide
4NPD = 4-nitro-o-phenylendiamine
9AA = 9-aminoacridine
2AA = 2-Amino-anthracene
C = Contamination, only 2 individual values
Table 4. 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 |
|||||
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
||
– |
Negative control |
13.3 ± 1.5 |
117.7 ± 10.0 |
7.7 ± 2.9 |
9.0 ± 4.4 |
33.0 ± 1.0 |
– |
Solvent control |
10.3 ± 5.8 |
111.7 ± 5.0 |
7.3 ± 0.6 |
9.3 ± 1.5 |
30.0 ± 6.3 |
– |
8 |
11.3 ± 2.3 |
118.3 ± 12.5 |
7.3 ± 1.5 |
13.0 ± 2.0 |
32.0 ± 12.5 |
– |
40 |
14.0 ± 2.7 |
105.0 ± 17.4 |
9.3 ± 0.6 |
9.3 ± 0.6 |
25.0 ± 6.2 |
– |
200 |
13.0 ± 1.0 |
97.7 ± 2.9 |
8.0 ± 1.7 |
7.7 ± 2.1 |
32.0 ± 6.6 |
– |
1000 |
13.3 ± 2.1 |
111.7 ± 9.0 |
9.7 ± 4.0M |
8.7 ± 4.0 |
41.3 ± 4.2 |
– |
5000 |
12.0 ± 1.0 |
125.3 ± 4.9 |
7.3 ± 2.1 |
11.0 ± 4.6 |
30.7 ± 2.5 |
Positive controls, –S9 |
Name |
SA |
SA |
9AA |
4NPD |
4NPD |
Concentrations (μg/plate) |
2 |
2 |
80 |
40 |
40 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
807.3 ±10.6 |
945.3 ± 37.5 |
1156.0 ± 216.5 |
2159.3 ± 106.6 |
226.7 ± 24.7 |
|
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
|
Concentrations (μg/plate) |
2.5 |
5 |
2.5 |
5 |
5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
12.3 ± 5.5 |
110.3 ± 13.1 |
9.3 ± 3.8 |
22.0 ± 6.1 |
29.3 ± 3.8 |
|
+ |
Negative control |
12.3 ± 5.8 |
121.7 ± 10.3 |
9.3 ± 2.3 |
14.0 ± 7.2 |
39.7 ± 3.1 |
+ |
Solvent control |
12.3 ± 3.1 |
120.3 ± 17.0 |
8.7 ± 1.2 |
11.3 ± 1.5 |
45.0 ± 1.0 |
+ |
8 |
15.0 ± 3.5 |
122.3 ± 12.7 |
6.7 ± 2.5 |
9.7 ± 1.2 |
40.1 ± 2.3 |
+ |
40 |
13.3 ± 2.3 |
123.0 ± 7.6 |
8.7 ± 5.0 |
13.7 ± 6.7 |
41.3 ± 8.3 |
+ |
200 |
16.0 ± 3.0 |
122.3 ± 13.3 |
9.0 ± 3.6 |
9.7 ± 6.4 |
45.0 ± 10.2 |
+ |
1000 |
16.0 ± 4.6 |
104.0 ± 19.3 |
8.3 ± 1.2 |
12.3 ± 3.2 |
41.7 ± 6.4 |
+ |
5000 |
16.0 ± 1.7 |
95.0 ± 8.5 |
11.7 ± 3.5 |
12.0 ± 5.6 |
39.0 ± 13.1 |
Positive controls, +S9 |
Name |
SA |
SA |
9AA |
4NPD |
4NPD |
Concentrations (μg/plate) |
2 |
2 |
80 |
40 |
40 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
232.0 ± 15.1 |
286.0 ± 29.2 |
748.0 ± 139.2 |
455.3 ± 14.2 |
531.3 ± 22.7 |
|
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
|
Concentrations (μg/plate) |
2.5 |
5 |
2.5 |
5 |
5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
131.0 ± 5.2 |
1735.0 ± 84.5 |
81.3 ± 8.4 |
585.3 ± 32.1 |
594.3 ± 56.6 |
|
SA = sodium azide
4NPD = 4-nitro-o-phenylendiamine
9AA = 9-aminoacridine
2AA = 2-Amino-anthracene
M = Manual evaluation
COMPARISON WITH HISTORICAL DATA
Table 1. Characteristic spontaneous mutation range of the test batches without S9-mix.
Tester strains |
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
Historical laboratory values (Mean values) Extreme values |
6 1-25 |
87 35-201 |
7 1-24 |
15 6-27 |
20 5-39 |
Ames et al.* (Mean values) Extreme values |
20 5-50 |
160 60-220 |
7 3-25 |
25 5-40 |
40 15-75 |
*Ames, BN, McCann, J, Yamasaki, E 1977, ´Methods for detecting carcinogens and mutagens with the Salmonella/mammalian microsome mutagenicity test.`in Kilbey et al. Handbook of Mutagenicity Test Procedures, Elsevier, Amsterdam, pp. 1 -17.
Table 2. Characteristic spontaneous mutation range of the test batches containing S9-mix.
Tester strains |
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
Historical laboratory values (Mean values) Extreme values |
8 1-25 |
105 54-252 |
6 1-23 |
18 3-48 |
27 7-76 |
STUDY RESULTS
Table 3. 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 |
|||||
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
||
– |
Negative control |
9.7 ± 3.1 |
97.7 ± 2.1 |
6.3 ± 3.2 |
6.7 ± 1.2 |
46.7 ± 3.2 |
– |
Solvent control |
7.7 ± 3.5 |
94.3 ± 17.4 |
8.0 ± 1.7 |
7.3 ± 2.3 |
46.3 ± 6.0 |
– |
8 |
8.3 ± 2.1 |
70.0 ± 7.8 |
9.0 ± 1.0 |
6.7 ± 1.2 |
33.7 ± 0.6 |
– |
40 |
11.0 ± 2.7 |
74.3 ± 3.2 |
7.3 ± 2.1 |
5.3 ± 0.6 |
40.3 ± 4.9 |
– |
200 |
12.0 ± 4.6 |
67.3 ± 11.2 |
6.3 ± 1.2 |
5.7 ± 1.2 |
37.3 ± 3.2 |
– |
1000 |
10.0 ± 4.0 |
62.7 ± 10.8 |
10.3 ± 1.2 |
4.3 ± 1.5 |
42.7 ± 3.1 |
– |
5000 |
9.0 ± 1.7 |
83.0 ± 6.9 |
5.7 ± 1.5 |
4.3 ± 0.6 |
48.3 ± 3.5 |
Positive controls, –S9 |
Name |
SA |
SA |
9AA |
4NPD |
4NPD |
Concentrations (μg/plate) |
2 |
2 |
80 |
40 |
40 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
744.7 ± 49.3 |
894.0 ± 62.0 |
939.7 ± 181.4 |
1541.0 ± 96.0 |
1410.7 ± 154.4 |
|
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
|
Concentrations (μg/plate) |
2.5 |
5 |
2.5 |
5 |
5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
9.0 ± 1.7 |
88.7 ± 17.7 |
7.0 ± 2.7 |
10.3 ± 1.5 |
46.0 ± 8.7 |
|
+ |
Negative control |
5.7 ± 0.6 |
80.3 ± 15.3 |
8.7 ± 2.9 |
11.3 ± 2.3 |
53.3 ± 3.2 |
+ |
Solvent control |
9.7 ± 4.6 |
80.3 ± 14.4 |
9.3 ± 1.2 |
13.7 ± 2.3 |
44.3 ± 6.7 |
+ |
8 |
12.3 ± 3.1 |
82.3 ± 12.1 |
7.7 ± 3.5 |
14.7 ± 2.9 |
49.7 ± 4.2 |
+ |
40 |
11.3 ± 4.6 |
92.0 ± 11.8 |
6.3 ± 0.6 |
13.0 ± 4.2 |
47.7 ± 4.0 |
+ |
200 |
8.0 ± 2.0 |
84.3 ± 4.9 |
8.0 ± 1.0 |
16.3 ± 1.5 |
53.3 ± 4.0 |
+ |
1000 |
10.0 ± 2.0 |
70.7 ± 9.7 |
9.3 ± 3.8 |
13.0 ± 1.0 |
50.5 ± 3.5C |
+ |
5000 |
10.0 ± 3.6 |
74.7 ± 11.0 |
7.0 ± 1.7 |
13.0 ± 3.0 |
54.0 ± 7.0 |
Positive controls, +S9 |
Name |
SA |
SA |
9AA |
4NPD |
4NPD |
Concentrations (μg/plate) |
2 |
2 |
80 |
40 |
40 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
213.3 ± 19.8 |
230.0 ± 4.6 |
283.3 ± 27.8 |
1434.0 ± 68.9 |
1307.6 ± 68.3 |
|
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
|
Concentrations (μg/plate) |
2.5 |
5 |
2.5 |
5 |
5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
186.3 ± 19.4 |
1328.3 ± 166.5 |
52.3 ± 1.5 |
1711.3 ± 25.0 |
1587.7 ± 237.7 |
|
SA = sodium azide
4NPD = 4-nitro-o-phenylendiamine
9AA = 9-aminoacridine
2AA = 2-Amino-anthracene
C = Contamination, only 2 individual values
Table 4. 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 |
|||||
TA1535 |
TA100 |
TA1537 |
TA1538 |
TA98 |
||
– |
Negative control |
13.3 ± 1.5 |
117.7 ± 10.0 |
7.7 ± 2.9 |
9.0 ± 4.4 |
33.0 ± 1.0 |
– |
Solvent control |
10.3 ± 5.8 |
111.7 ± 5.0 |
7.3 ± 0.6 |
9.3 ± 1.5 |
30.0 ± 6.3 |
– |
8 |
11.3 ± 2.3 |
118.3 ± 12.5 |
7.3 ± 1.5 |
13.0 ± 2.0 |
32.0 ± 12.5 |
– |
40 |
14.0 ± 2.7 |
105.0 ± 17.4 |
9.3 ± 0.6 |
9.3 ± 0.6 |
25.0 ± 6.2 |
– |
200 |
13.0 ± 1.0 |
97.7 ± 2.9 |
8.0 ± 1.7 |
7.7 ± 2.1 |
32.0 ± 6.6 |
– |
1000 |
13.3 ± 2.1 |
111.7 ± 9.0 |
9.7 ± 4.0M |
8.7 ± 4.0 |
41.3 ± 4.2 |
– |
5000 |
12.0 ± 1.0 |
125.3 ± 4.9 |
7.3 ± 2.1 |
11.0 ± 4.6 |
30.7 ± 2.5 |
Positive controls, –S9 |
Name |
SA |
SA |
9AA |
4NPD |
4NPD |
Concentrations (μg/plate) |
2 |
2 |
80 |
40 |
40 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
807.3 ±10.6 |
945.3 ± 37.5 |
1156.0 ± 216.5 |
2159.3 ± 106.6 |
226.7 ± 24.7 |
|
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
|
Concentrations (μg/plate) |
2.5 |
5 |
2.5 |
5 |
5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
12.3 ± 5.5 |
110.3 ± 13.1 |
9.3 ± 3.8 |
22.0 ± 6.1 |
29.3 ± 3.8 |
|
+ |
Negative control |
12.3 ± 5.8 |
121.7 ± 10.3 |
9.3 ± 2.3 |
14.0 ± 7.2 |
39.7 ± 3.1 |
+ |
Solvent control |
12.3 ± 3.1 |
120.3 ± 17.0 |
8.7 ± 1.2 |
11.3 ± 1.5 |
45.0 ± 1.0 |
+ |
8 |
15.0 ± 3.5 |
122.3 ± 12.7 |
6.7 ± 2.5 |
9.7 ± 1.2 |
40.1 ± 2.3 |
+ |
40 |
13.3 ± 2.3 |
123.0 ± 7.6 |
8.7 ± 5.0 |
13.7 ± 6.7 |
41.3 ± 8.3 |
+ |
200 |
16.0 ± 3.0 |
122.3 ± 13.3 |
9.0 ± 3.6 |
9.7 ± 6.4 |
45.0 ± 10.2 |
+ |
1000 |
16.0 ± 4.6 |
104.0 ± 19.3 |
8.3 ± 1.2 |
12.3 ± 3.2 |
41.7 ± 6.4 |
+ |
5000 |
16.0 ± 1.7 |
95.0 ± 8.5 |
11.7 ± 3.5 |
12.0 ± 5.6 |
39.0 ± 13.1 |
Positive controls, +S9 |
Name |
SA |
SA |
9AA |
4NPD |
4NPD |
Concentrations (μg/plate) |
2 |
2 |
80 |
40 |
40 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
232.0 ± 15.1 |
286.0 ± 29.2 |
748.0 ± 139.2 |
455.3 ± 14.2 |
531.3 ± 22.7 |
|
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
|
Concentrations (μg/plate) |
2.5 |
5 |
2.5 |
5 |
5 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
131.0 ± 5.2 |
1735.0 ± 84.5 |
81.3 ± 8.4 |
585.3 ± 32.1 |
594.3 ± 56.6 |
|
SA = sodium azide
4NPD = 4-nitro-o-phenylendiamine
9AA = 9-aminoacridine
2AA = 2-Amino-anthracene
M = Manual evaluation
Table 1. Summary of data obtained in experiment #1a.
Test item |
Concentration |
Mitotic Index |
Aberrant cells in % |
|
|
in µg/mL |
in % mean |
with gaps |
without gaps |
Exposure period 18h, without S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
10.7 |
2.0 |
0.0 |
MMC |
0.03 |
2.1 |
24.6 |
18.5* |
MMC |
0.04 |
2.9 |
40.0 |
31.9* |
Test substance |
10 |
8.0 |
3.5 |
1.5 |
40 |
6.7 |
3.0 |
0.5 |
|
80 |
5.3 |
5.0 |
0.0 |
|
Exposure period 18h, with S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
7.0 |
6.5 |
4.0 |
CP |
3.0 |
2.7 |
45.0 |
34.0* |
Test substance |
10 |
6.7 |
3.5 |
1.5 |
60 |
6.1 |
4.5 |
1.0 |
|
100 |
5.7 |
4.0 |
2.0 |
|
MMC: Mitomycin C; CP: Cyclophosphamide (positive controls)
*: significant, no statistical evaluation
Table 2. Summary of data obtained in experiment #2.
Test item |
Concentration |
Mitotic Index |
Aberrant cells in % |
|
|
in µg/mL |
in % mean |
with gaps |
without gaps |
Exposure period 18h, without S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
7.9 |
6.0 |
3.5 |
MMC |
0.03 |
3.4 |
23.5 |
14.0* |
Test substance |
10 |
7.5 |
4.0 |
0.5 |
40 |
8.0 |
9.5 |
3.0 |
|
80 |
5.8 |
5.5 |
0.5 |
|
Exposure period 18h, with S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
6.3 |
8.0 |
2.0 |
CP |
3.0 |
1.6 |
42.0 |
33.3* |
Test substance |
10 |
6.5 |
5.5 |
0.0 |
60 |
5.7 |
4.5 |
1.5 |
|
100 |
6.7 |
6.5 |
1.5 |
|
Exposure period 28h, without S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
7.9 |
3.5 |
0.0 |
MMC |
0.03 |
4.4 |
43.5 |
32.5* |
Test substance |
80 |
5.8 |
4.5 |
0.5 |
Exposure period 28h, with S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
9.2 |
6.5 |
1.5 |
CP |
3.0 |
6.0 |
36.5 |
27.5* |
Test substance |
100 |
8.4 |
3.5 |
1.0 |
MMC: Mitomycin C; CP: Cyclophosphamide (positive controls)
*: significant, no statistical evaluation
Table 1. Summary of data obtained in experiment #1a.
Test item |
Concentration |
Mitotic Index |
Aberrant cells in % |
|
|
in µg/mL |
in % mean |
with gaps |
without gaps |
Exposure period 18h, without S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
10.7 |
2.0 |
0.0 |
MMC |
0.03 |
2.1 |
24.6 |
18.5* |
MMC |
0.04 |
2.9 |
40.0 |
31.9* |
Test substance |
10 |
8.0 |
3.5 |
1.5 |
40 |
6.7 |
3.0 |
0.5 |
|
80 |
5.3 |
5.0 |
0.0 |
|
Exposure period 18h, with S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
7.0 |
6.5 |
4.0 |
CP |
3.0 |
2.7 |
45.0 |
34.0* |
Test substance |
10 |
6.7 |
3.5 |
1.5 |
60 |
6.1 |
4.5 |
1.0 |
|
100 |
5.7 |
4.0 |
2.0 |
|
MMC: Mitomycin C; CP: Cyclophosphamide (positive controls)
*: significant, no statistical evaluation
Table 2. Summary of data obtained in experiment #2.
Test item |
Concentration |
Mitotic Index |
Aberrant cells in % |
|
|
in µg/mL |
in % mean |
with gaps |
without gaps |
Exposure period 18h, without S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
7.9 |
6.0 |
3.5 |
MMC |
0.03 |
3.4 |
23.5 |
14.0* |
Test substance |
10 |
7.5 |
4.0 |
0.5 |
40 |
8.0 |
9.5 |
3.0 |
|
80 |
5.8 |
5.5 |
0.5 |
|
Exposure period 18h, with S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
6.3 |
8.0 |
2.0 |
CP |
3.0 |
1.6 |
42.0 |
33.3* |
Test substance |
10 |
6.5 |
5.5 |
0.0 |
60 |
5.7 |
4.5 |
1.5 |
|
100 |
6.7 |
6.5 |
1.5 |
|
Exposure period 28h, without S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
7.9 |
3.5 |
0.0 |
MMC |
0.03 |
4.4 |
43.5 |
32.5* |
Test substance |
80 |
5.8 |
4.5 |
0.5 |
Exposure period 28h, with S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
9.2 |
6.5 |
1.5 |
CP |
3.0 |
6.0 |
36.5 |
27.5* |
Test substance |
100 |
8.4 |
3.5 |
1.0 |
MMC: Mitomycin C; CP: Cyclophosphamide (positive controls)
*: significant, no statistical evaluation
Table 1: Cytotoxic and mutagenic responses
Treatment |
Concentration [µg/mL] |
Cloning efficiency [%] |
Relative total growth [%] |
Mutation frequency x 10-6
|
||
total |
small colonies |
large colonies |
||||
3 h treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
86 |
100 |
61 |
42 |
19 |
Test substance |
0.1 |
101 |
114 |
56 |
29 |
25 |
0.3 |
101 |
108 |
52 |
28 |
23 |
|
1.0 |
118 |
140 |
66 |
48 |
16 |
|
3.0 |
91 |
109 |
63 |
46 |
15 |
|
10.0 |
95 |
108 |
66 |
32 |
33 |
|
33.0 |
97 |
104 |
76 |
46 |
28 |
|
100.0* |
91 |
97 |
81 |
32 |
46 |
|
333.0* |
102 |
77 |
64 |
48 |
14 |
|
MMS |
15 |
60 |
45 |
685 |
521 |
118 |
3 h treatment with 8% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
91 |
100 |
67 |
36 |
29 |
Test substance |
0.1 |
108 |
111 |
74 |
47 |
25 |
0.3 |
89 |
94 |
71 |
45 |
24 |
|
1.0 |
108 |
113 |
64 |
42 |
20 |
|
3.0 |
98 |
107 |
78 |
53 |
23 |
|
10.0 |
95 |
100 |
87 |
54 |
29 |
|
33.0 |
108 |
96 |
55 |
32 |
22 |
|
100.0* |
98 |
89 |
83 |
60 |
21 |
|
333.0* |
94 |
92 |
63 |
23 |
39 |
|
CP |
7.5 |
60 |
32 |
1074 |
829 |
144 |
24 h treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
115 |
100 |
51 |
26 |
23 |
Test substance |
3 |
135 |
92 |
58 |
42 |
14 |
10 |
137 |
109 |
51 |
39 |
11 |
|
33 |
133 |
85 |
71 |
32 |
35 |
|
100* |
110 |
30 |
100 |
35 |
60 |
|
125* |
118 |
31 |
70 |
31 |
36 |
|
140* |
118 |
20 |
103 |
51 |
47 |
|
175* |
102 |
9 |
134 |
53 |
72 |
|
MMS |
5 |
101 |
73 |
865 |
463 |
233 |
3 h treatment with 12% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
109 |
100 |
72 |
47 |
23 |
Test substance |
0.1 |
110 |
100 |
73 |
49 |
21 |
0.3 |
123 |
117 |
72 |
47 |
23 |
|
1.0 |
104 |
105 |
82 |
52 |
27 |
|
3.0 |
97 |
104 |
94 |
62 |
29 |
|
10.0 |
115 |
126 |
87 |
57 |
26 |
|
33.0 |
107 |
108 |
85 |
59 |
23 |
|
100.0* |
131 |
123 |
80 |
51 |
26 |
|
333.0* |
97 |
83 |
92 |
64 |
24 |
|
CP |
7.5 |
70 |
59 |
979 |
621 |
221 |
*precipitation of test substance in the exposure medium
MMS = methylmethanesulfonate
CP = cyclophosphamide
Table 1: Cytotoxic and mutagenic responses
Treatment |
Concentration [µg/mL] |
Cloning efficiency [%] |
Relative total growth [%] |
Mutation frequency x 10-6
|
||
total |
small colonies |
large colonies |
||||
3 h treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
86 |
100 |
61 |
42 |
19 |
Test substance |
0.1 |
101 |
114 |
56 |
29 |
25 |
0.3 |
101 |
108 |
52 |
28 |
23 |
|
1.0 |
118 |
140 |
66 |
48 |
16 |
|
3.0 |
91 |
109 |
63 |
46 |
15 |
|
10.0 |
95 |
108 |
66 |
32 |
33 |
|
33.0 |
97 |
104 |
76 |
46 |
28 |
|
100.0* |
91 |
97 |
81 |
32 |
46 |
|
333.0* |
102 |
77 |
64 |
48 |
14 |
|
MMS |
15 |
60 |
45 |
685 |
521 |
118 |
3 h treatment with 8% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
91 |
100 |
67 |
36 |
29 |
Test substance |
0.1 |
108 |
111 |
74 |
47 |
25 |
0.3 |
89 |
94 |
71 |
45 |
24 |
|
1.0 |
108 |
113 |
64 |
42 |
20 |
|
3.0 |
98 |
107 |
78 |
53 |
23 |
|
10.0 |
95 |
100 |
87 |
54 |
29 |
|
33.0 |
108 |
96 |
55 |
32 |
22 |
|
100.0* |
98 |
89 |
83 |
60 |
21 |
|
333.0* |
94 |
92 |
63 |
23 |
39 |
|
CP |
7.5 |
60 |
32 |
1074 |
829 |
144 |
24 h treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
115 |
100 |
51 |
26 |
23 |
Test substance |
3 |
135 |
92 |
58 |
42 |
14 |
10 |
137 |
109 |
51 |
39 |
11 |
|
33 |
133 |
85 |
71 |
32 |
35 |
|
100* |
110 |
30 |
100 |
35 |
60 |
|
125* |
118 |
31 |
70 |
31 |
36 |
|
140* |
118 |
20 |
103 |
51 |
47 |
|
175* |
102 |
9 |
134 |
53 |
72 |
|
MMS |
5 |
101 |
73 |
865 |
463 |
233 |
3 h treatment with 12% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
109 |
100 |
72 |
47 |
23 |
Test substance |
0.1 |
110 |
100 |
73 |
49 |
21 |
0.3 |
123 |
117 |
72 |
47 |
23 |
|
1.0 |
104 |
105 |
82 |
52 |
27 |
|
3.0 |
97 |
104 |
94 |
62 |
29 |
|
10.0 |
115 |
126 |
87 |
57 |
26 |
|
33.0 |
107 |
108 |
85 |
59 |
23 |
|
100.0* |
131 |
123 |
80 |
51 |
26 |
|
333.0* |
97 |
83 |
92 |
64 |
24 |
|
CP |
7.5 |
70 |
59 |
979 |
621 |
221 |
*precipitation of test substance in the exposure medium
MMS = methylmethanesulfonate
CP = cyclophosphamide
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
There are no data for genetic toxicity available for the target substance. In accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5 read-across from appropriate substances is conducted to fulfill the standard information requirements set out in Regulation (EC) No 1907/2006, Annex VIII, 8.4.
According to Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met”. In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across) “to avoid the need to test every substance for every endpoint”.
The target substance Fatty acids, C16-18 and C18-unsatd., esters with propylene glycol represents a UVCB substance predominantly comprised of diesters of an aliphatic diol (1,2-propyleneglycol (PG)) chemically linked to mainly oleic acid (C18:1) but as well to palmitic acid (C16), palmitoleic acid (C16:1), stearic acid (C18) and/or linoleic acid (C18:2).
Glycol esters are in general known to be stepwise hydrolysed by gastrointestinal enzymes into the free fatty acid component and the respective alcohol (Long, 1958; Lehninger, 1970; Mattson and Volpenhein, 1972).
Based on the common metabolic fate of glycol esters, the read-across approach is based on the presence of common functional groups, common precursors and the likelihood of common breakdown products via biological processes, which result in structurally similar chemicals, common functional groups, structural similarities and similar physico-chemical, toxicological and toxicokinetic behaviour. For further details on the read-across approach, please refer to the analogue justification in section 13 of the technical dossier.
As no data are available on mutagenic properties of the target substance, read-across to reliable data on the analogue substances Fatty acids, C14-18 and C16-18-unsatd., esters with propylene glycol (CAS 84988-75-0), C8-C10-1,3-Butandiolester (CAS 853947-59-8) and Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1) was conducted.
Genetic toxicity (mutagenicity) in bacteria in vitro
CAS 84988-75-0
A study investigating the genetic toxicity in vitro of Fatty acids, C14-18 and C16-18-unsatd., esters with propylene glycol is available. The study was conducted according to OECD guideline 471 under GLP conditions (Banduhn, 1991). In two independent experiments, the tester strains Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 were tested according to the plate incorporation procedure. Concentrations from 8 to 5000 µg/plate were investigated with and without a metabolic activation system (Aroclor 1254-induced rat liver S9-mix). No increase in the number of revertant colonies was noted in any of the bacterial strains, with and without metabolic activation system. No cytotoxicity was observed up to the highest dose tested. The included positive and negative controls showed the expected results and were therefore considered as valid. Thus, under the conditions of this study, the test substance did not induce mutations in the bacterial mutation tests in the absence and presence of a metabolic activation system in any of the strains tested.
Genetic toxicity (cytogenicity) in mammalian cells in vitro
CAS 853947-59-8
An in vitro mammalian chromosome aberration test was conducted with C8-C10-1,3-Butandiolester in accordance with OECD guideline 473 under GLP conditions (Dechert, 1997). The induction of structural chromosome aberrations was evaluated in vitro in Chinese hamster lung fibroblasts (V79 cells), incubated for 18 and 28 h with and without a metabolic activation system (S9-mix from rats treated with Aroclor 1245). Concentrations of 10-100 µg/mL (18 h incubation) and 80 and 100 µg/mL (28 h incubation) of the test substance in the vehicle ethanol were applied. The solubility limit of the test substance in the vehicle ethanol in the culture medium was determined to be 100 µg/mL. In the first experiment without metabolic activation, the negative controls exhibited a mitotic index of 2.0% only and the experiment was therefore repeated. Thereafter, the negative as well as the positive controls showed the expected results and were within the range of historical control data. The frequency of polyploidy cells with and without metabolic activation was within the expected range (< 10%). In the experiments both with and without metabolic activation, a systematic influence of the test substance was observed, which led to a reduction in the mitotic index. No statistically or biologically significant increase in the incidence of chromosome aberrations was observed.
Therefore, under the conditions of the study, C8-C10-1,3-Butandiolester did not show clastogenic activity in this chromosomal aberration test with and without metabolic activation performed in Chinese hamster lung fibroblasts in vitro.
Genetic toxicity (mutagenicity) in mammalian cells in vitro
CAS 91031-31-1
Mutagenic properties of Fatty acids, C16-18, esters with ethylene glycol were characterized in an in vitro mammalian cell gene mutation study according to OECD guideline 476 under GLP conditions (Verspeek-Rip, 2010). Gene mutations in the thymidine kinase locus were investigated in L5178Y mouse lymphoma cells in the presence and absence of a metabolic activation system (Phenobarbital/β-naphtoflavone-induced rat liver S9). In the first experiment, cells were exposed for 3 h to the test substance at concentrations of 0.1-333 µg/mL (in DMSO) with and without metabolic activation. Concentrations of the second experiment without metabolic activation for an exposure time of 24 h ranged from 3-175 µg/mL and with metabolic activation (3 h; 12% S9-mix) from 0.1-333 µg/mL. The vehicle and positive controls in the study showed the expected results and were within the range of historical control data. No cytotoxicity was observed up to the precipitating concentration of 100 µg/mL and up to 333 µg/mL, respectively. There was no significant increase in the number of forward mutations at the thymidine kinase locus of L5178Y mouse lymphoma cells treated with the test material, neither in the presence nor in the absence of a metabolic activation system. Under the conditions of the study, Fatty acids, C16-18, esters with ethylene glycol did not show gene mutation activity in this test performed in L5178Y mouse lymphoma cells in vitro.
Conclusion on genetic toxicity
The available data on read-across analogue substances do not provide evidence that Fatty acids, C14-18 and C16-18-unsatd., esters with propylene glycol (CAS 84988-75-0), C8-C10-1,3-Butandiolester (CAS 853947-59-8) or Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1) exhibit mutagenic or clastogenic properties in either bacteria or mammalian cells. Therefore, no properties for genetic toxicity are expected for the target substance.
References
Agency for Toxic Substances and Disease Registry (ATSDR) (1997). Toxicological Profile for Propylene Glycol. US Department of Health and Human Services. Atlanta, US.
Agency for Toxic Substances and Disease Registry (ATSDR) (2010). Toxicological Profile for Ethylene Glycol. US Department of Health and Human Services. Atlanta, US.
Lehninger, A.L. (1970). Biochemistry. Worth Publishers, Inc.Long, C.L. et al. (1958). Studies on absorption and metabolism of propylene glycol distearate. Arch Biochem Biophys, 77(2):428-439.
Mattson, F.H. and Volpenhein, R.A. (1972). Hydrolysis of fully esterified alcohols containing from one to eight hydroxyl groups by the lipolytic enzymes of the rat pancreatic juice. Journal of Lipid Research 13: 325-328
Miller, O.N., Bazzano, G. (1965).Propanediol metabolism and its relation to lactic acid -metabolism. Annals of the New York Academy of Sciences 119:957-973.
Ritchie, A.D. (1927). Lactic acid in fish and crustacean muscle. Journal of Experimental Biology 4:327-332.
Justification for selection of genetic toxicity endpoint: Hazard assessment is conducted by means of read-across from structural analogues. All available studies are adequate and reliable based on the identified similarities in structure and intrinsic properties between source and target substances and overall quality assessment (refer to the endpoint discussion for further details). No study was selected, since all available in vitro genetic toxicity studies were negative. Short description of key information: Genetic toxicity in vitro: Ames test (OECD 471): negative with and without metabolic activation in S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 Chromosome aberration (OECD 473): negative with and without metabolic activation in Chinese hamster lung fibroblasts (V79) cells Gene mutation in mammalian cells (OECD 476): CAS 91031-31-1: negative with and without metabolic activation in L5178Y mouse lymphoma cells Endpoint Conclusion: No adverse effect observed (negative).
Justification for classification or non-classification
Based on the analogue read-across approach, the available data on genetic toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification.
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.
