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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: - | CAS number: -
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Negative results in Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537 and E.coli with and without metabolic activation (OECD 471, GLP).
Negative results in mammalian cell gene mutation tests using mouse lymphoma cells, with and without metabolic activation (OECD 476, GLP).
Negative results in mammalian cytogenicity testing with human peripheral blood lymphocytes (OECD 473, GLP).
Read across form the source substance oleic acid, monoester with oxybis(propanediol), (CAS 49553-76-6)
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:
- key study
- Study period:
- 14 - 24 Sep 2012
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP - Guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his operon (for S. typhimurium) and trp operon (for E. coli)
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Metabolic activation system:
- co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254.
- Test concentrations with justification for top dose:
- Preliminary cytotoxicity test (Experiment I): 33.3, 66.7, 100, 333, 667, 1000, 3333, and 5000 µg/plate with and without metabolic activation
Main assay (Experiment II): 333, 667, 1000, 3333, and 5000 µg/plate with and without metabolic activation - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: DMSO was chosen as the dosing vehicle based on the solubility of the test substance and compatibility with the target cells. The test substance was soluble in DMSO at 50 mg/mL, the highest stock concentration that was prepared for use on this study. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- -S9: 2-NF (1 µg/plate, TA 98), SA (2 µg/plate, TA 100 and TA 1535), ICR-191 (2 µg/plate, TA 1537), 4-NQO (1 µg/plate, WP2 uvrA); +S9: BaP (2.5 µg/plate, TA 98); 2-AA (2.5 µg/plate, TA 100, TA 1535 and TA 1537; 25 µg/plate, WP2 uvrA)
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 2-nitrofluorene
- sodium azide
- benzo(a)pyrene
- other: acridine mutagen ICR-191 and 2-aminoanthracene
- Remarks:
- BaP: benzo[a]pyrene; 4-NQO: 4-nitroquinoline N-oxide; acridine mutagen ICR-191: ICR-191; SA: sodium azide; 2-aminoanthracene: 2-AA; 2-NF: 2-nitrofluorene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48-72 h
NUMBER OF REPLICATIONS: duplicate plates in the preliminary cytotoxicity test and triplicate plates in the main assay
DETERMINATION OF CYTOTOXICITY
- Method: mean number of revertant colonies, inspection of bacterial background lawn - Evaluation criteria:
- Criteria for a positive response:
- Strains TA1535 and TA1537: data were judged positive if the increase in mean revertants at the highest numerical dose response was ≥ 3.0-fold the mean concurrent negative control value (vehicle control). This increase in the mean number of revertants per plate had to be accompanied by a dose response associated with increasing concentrations of the test substance unless observed at the top dose level only.
- Strains TA98, TA100 and WP2uvrA: data sets were judged positive if the increase in mean revertants at the highest numerical dose response was ≥ 2.0-fold the mean concurrent negative control value (vehicle control). This increase in the mean number of revertants per plate has to be accompanied by a dose response associated with increasing concentrations of the test substance unless observed at the top dose level only. - Statistics:
- For each tester strain, the mean of the number of revertants and the standard deviations were calculated.
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 5000 µg/plate: TA 1535 (+S9)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 5000 µg/plate: TA 1537 (-S9)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: in the absence of S9 mix, test substance precipitation was observed starting at 3333 μg/plate for all tester strains with the exception of WP2 uvrA where precipitation was observed only at 5000 μg/plate. In the presence of S9 mix, precipitation was observed starting at 3333 μg/plate for all tester strains with the exception of TA 100 and WP2uvrA. Precipitation was not observed with WP2 uvrA in the activated condition and was only observed at 5000 μg/plate with tester strain TA 100. However, none of the precipitates prevented accurate colony counting.
RANGE-FINDING/SCREENING STUDIES: in the preliminary cytotoxicity test, a 50% reduction in revertant colonies was observed with TA 1537 in the presence of metabolic activation starting at 3333 µg/plate. No cytotoxicity was observed in any of the other tester strains after treatment with concentrations ranging from 33.3 to 5000 µg/plate in the presence and absence of metabolic activation, respectively.
COMPARISON WITH HISTORICAL CONTROL DATA: the mean revertant number of the vehicle and positive controls was within their respective historical ranges.
ADDITIONAL INFORMATION ON CYTOTOXICITY: a >50% reduction in revertant colonies was observed at 5000 μg/plate for both TA1537 in the absence of S9 activation, as well as for TA1535 in the presence of S9 activation. - 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:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 16 Aug - 12 Oct 2012
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP - Guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5375 - In vitro Mammalian Chromosome Aberration Test
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- not applicable
- Species / strain / cell type:
- other: human peripheral blood lymphocytes (from healthy donor(s) less than 50 years old without previous chemotherapy or radiotherapy; and without recent (within the last 6 months) viral disease or X-ray exposure)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: complete medium (RPMI 1640 medium containing approximately 15% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units penicillin/mL, and 100 μg streptomycin/mL) supplemented with 1-2% phytohemagglutinin-M (PHA-M)
- Properly maintained: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254
- Test concentrations with justification for top dose:
- Without S9-mix, 4-h exposure: 25, 50, 100, 150, 200 µg/mL
With S9-mix, 4-h exposure: 50, 100, 200, 300, 400 µg/mL
Without S9-mix, 22-h exposure: 10, 25, 50, 75, 100 µg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The vehicle of choice for this study was DMSO, which permitted preparation of the highest workable/soluble stock concentration. Under the conditions of this test system, the final concentration of DMSO in the treatment medium did not exceed 1% of the treatment medium. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- Remarks:
- mitomycin C (-S9): 0.2 µg/mL (4 and 22 h exposure period), cyclophosphamide (+S9): 10 µg/mL for the 4 h exposure period
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 and 22 h
- Fixation time (start of exposure up to fixation or harvest of cells): 22 h
SPINDLE INHIBITOR (cytogenetic assays): colcemid (0.1 µg/mL medium)
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: duplicate cultures
NUMBER OF CELLS EVALUATED: 100 from each duplicate culture
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreplication: yes - Evaluation criteria:
- An assay was considered acceptable for evaluation of test results only if all of the following criteria were satisfied.
Negative Controls: The frequency of cells with structural chromosome aberrations was within the negative historical control range.
Positive Controls: The percentage of cells with structural chromosome aberrations were statistically significantly greater (p < 0.05, Fisher’s exact test) than the vehicle control response.
The following conditions were used as a guide to determine a positive response:
• A statistically significant increase (p < 0.05, Fisher’s exact test) in the percentage of cells with structural aberrations was seen in one or more treatment groups relative to the vehicle control response.
• The observed increased frequencies were accompanied by a concentration-related increase.
• A statistically significant increase was observed at the highest dose only.
• Note: Statistically significant values that did not exceed the historical control range for the negative/vehicle control may be judged as not being biologically significant.
The following condition was used as a guide to determine an equivocal response:
• Results observed in any of the assays resulted in statistically significant elevations in structural chromosome aberrations at more than one test concentration level, except the highest dose, without demonstrating a dose-responsive trend.
The test substance was judged negative if the following condition was met:
• There was no statistically significant increase in the percentage of cells with structural aberrations in any treatment group relative to the vehicle control group. - Statistics:
- Data were evaluated using scientific judgment. Statistical analysis was used as a guide to determine whether or not the test substance induced a positive response. Statistical analysis consisted of a Fisher’s exact test (with Bonferroni-Holm Adjustment) to compare the percentage of cells with structural or numerical aberrations (or the percentage of cells with more than one aberration, if required) in the test substance treated groups with the vehicle control response. A Cochran-Armitage test for dose responsiveness was conducted only on values within a test condition only if statistically significant values, based on the Fisher’s exact test, are found. At the discretion of the study director, statistical analyses may be conducted on the percentage of cells with numerical aberrations as well.
- Species / strain:
- other: human peripheral blood lymphocytes
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 150 μg/mL in the 4-hour nonactivated test condition, at 100 μg/mL in the 22-hour non-activated test and in the 4-hour S9-activated test at 200 µg/mL and above
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH and osmolality: In the preliminary toxicity assays, osmolality and pH measurements were taken from 2 test substance concentrations (2500 and 5000 μg/mL) and the vehicle control media. The pH of the non-activated test system was 8.02 in the vehicle compared with 7.89 at 2500 μg/mL and 7.87 at 5000 μg/mL. The pH of the S9-activated test system was 7.50 in the vehicle compared with 7.37 at 2500 μg/mL and 7.34 at 5000 μg/mL. The osmolality of the nonactivated test system was 443 in the vehicle compared with 339 at 2500 μg/mL and 358 at 5000 μg/mL. The osmolality of the S9-activated test system was 462 in the vehicle compared with 418 at 2500 μg/mL and 384 at 5000 μg/mL. There were no observed increases in osmolality that were ≥ 20%, and therefore, not considered significant.
- Precipitation: Precipitation was observed in the 4-hour S9-activated test condition at the end of treatment only at 400 μg/mL.
RANGE-FINDING/SCREENING STUDIES: In the preliminary toxicity assay, the cells were treated for 4 and 22 hours in the non-activated test condition and for 4 hours in the S9-activated test condition. All cells were harvested 22 hours after treatment initiation. The cells were exposed to 9 concentrations of the test substance ranging from 25 to 5000 μg/mL, as well as a vehicle control. Precipitation was observed in the treatment media at ≥ 2500 μg/mL at the beginning of treatment in each test condition. In the 4-hour test conditions precipitation was observed at ≥ 500 μg/mL at the end of treatment while no precipitation was observed in the 22-hour test condition at the end of treatment. Substantial toxicity (greater than 50% reduction in mitotic index relative to the vehicle control) was observed at 250 μg/mL in the 4-hour non-activated test condition, at 500 μg/ml in the 4-hour S9-activated test condition and at 100 μg/mL in the 22-hour non-activated test condition. Based on the findings from the preliminary toxicity assay, the highest concentration chosen for the chromosome aberration assay was based on test substance induced toxicity.
COMPARISON WITH HISTORICAL CONTROL DATA:
Historical control data were given in the study report for the negative and positive controls of studies conducted from 2006 to 2011. The results for the positive and negative controls were within the historical control ranges given. - 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 gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 31 Aug - 08 Oct 2012
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP - Guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Office of Food Additive Safety, Redbook 2000
- 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: treatment medium: Fischer's Medium for Leukemic Cells of Mice with 0.1% Pluronics (F0P); restrictive medium for cleansing; washing medium: F0P supplemented with 10% horse serum, 2 mM L-glutamine, 100 U penicillin/mL and 100 μg streptomycin/mL (F10P); cloning medium: cloning medium (C.M.) containing 0.24% dissolved Noble agar in F0P plus 20% horse serum
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254
- Test concentrations with justification for top dose:
- Preliminary experiment: 0.5, 1.5, 5, 15, 50, 150, 1500 and 5000 µg/mL
- 4 h treatment: with and without metabolic activation
- 24 treatment: without metabolic activation
Main assay - concentrations used for treatment:
- Experiment I - 4 h treatment: 10 to 150 µg/mL with and without metabolic activation
- Experiment II -24 h treatment: 5 to 75 µg/mL without metabolic activation
Main assay - concentrations used for cloning:
- Experiment I - 4 h treatment: 10, 20, 30, 40 and 50 µg/mL with and without metabolic activation
- Experiment II -24 h treatment: 5, 10, 20, 30 and 40 µg/mL without metabolic activation - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: DMSO was selected as the solvent of choice based on the solubility of the test article and compatibility with the target cells. The test article formed a clear solution in DMSO at approximately 500 mg/mL, the maximum concentration tested in the solubility test. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- -S9: methylmethanesulfonate, 15 and 20 µg/mL (first experiment) and 5 and 7.5 µg/mL (second experiment); +S9: 7,12-Dimethyl-benz(a)anthracene, 1.25 and 1.5 µg/mL
- Positive control substance:
- 7,12-dimethylbenzanthracene
- methylmethanesulfonate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: Experiment I: 4 h (± S9); Experiment II: 24 h (-S9)
- Expression time (cells in growth medium): for expression of the mutant phenotype in the 4-hour exposure, the cultures were counted using an electronic cell counter and adjusted to 3E+5 cells/mL one and two days after treatment in 20 and 10 mL total volume, respectively. For the 24-hour exposure, cultures were adjusted to 3E+5 cells/mL in 20 mL immediately after test article removal, then two and three days after treatment in 20 and 10 mL total volume, respectively. For expression of the TK-/- cells, cells were placed in cloning medium (C.M.) containing 0.24% dissolved Noble agar in F0P plus 20% horse serum. Two flasks per culture to be cloned were labelled with the test article concentration, activation condition, and either TFT (trifluorothymidine, the selective agent) or VC (viable count). Each flask was filled with 100 mL C.M. and placed in an incubator shaker at 37 ± 1 °C until used. The cells were centrifuged at approximately 200-300 g for 10 minutes and the supernatant was decanted. The cells were then diluted in C.M. to concentrations of 3E+6 cells/100 mL C.M. for the TFT flask and 600 cells/100 mL C.M. for the VC flask. After the dilution, 1.0 mL of stock solution of TFT was added to the TFT flask (final concentration of 3 μg/mL) and both this flask and the VC flask were placed on the shaker at 125 rpm and 37 ± 1 °C. After 15 minutes, the flasks were removed and the cell suspension was dispensed equally into each of three appropriately labelled Petri dishes. To accelerate the gelling process, the plates were placed in cold storage (2-8 °C) for approximately 30 minutes. The plates were then incubated at 37 ± 1 °C in a humidified 5 ± 1% CO2 atmosphere for 10-14 days.
- Selection time (if incubation with a selection agent): 10-14 days
- Fixation time (start of exposure up to fixation or harvest of cells): 12-16 days
SELECTION AGENT (mutation assays): 3 μg/mL trifluorothymidine (TFT)
NUMBER OF REPLICATIONS: duplicate cultures each in two independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency; relative total growth; other: suspension growth, relative suspension growth
OTHER EXAMINATIONS:
- Other: small and large colonies were differentiated, as small colonies are capable to indicate chromosomal mutations.
OTHER: DETERMINATION OF OSMOLALITY: the osmolality of the solvent control and the highest soluble concentration in treatment medium were determined. - Evaluation criteria:
- In evaluation of the data, increases in induced mutant frequency that occurred only at highly toxic concentrations (i.e., less than 10% total growth) were not considered biologically relevant. All conclusions were based on scientific judgment; however, the following criteria are presented as a guide to interpretation of the data (Moore et al., 2006):
- a result was considered positive if a concentration-related increase in induced mutant frequency was observed in the treated cultures and one or more treatment conditions with 10% or greater total growth exhibited induced mutant frequencies of ≥ 90 mutants per 1E+6 clonable cells (based on the average mutant frequency of duplicate cultures). If the average solvent control mutant frequency was > 90 mutants per 1E+6 clonable cells, a doubling of mutant frequency over the background will also be required (Mitchell et al., 1997).
- a result was considered negative if the treated cultures exhibited induced mutant frequencies of less than 90 mutants per 1E+6 clonable cells (based on the average mutant frequency of duplicate cultures) and there was no concentration-related increase in mutant frequency.
- there are some situations in which a chemical would be considered negative when there was no culture showing between 10-20% survival: 1) There was no evidence of mutagenicity (e.g. no dose response or increase in induced mutant frequencies between 45 and 89 mutants per 106) in a series of data points within 100% to 20% survival and there was at least one negative data point between 20% and 25% survival. 2) There was no evidence of mutagenicity (e.g. no dose response or increase in induced mutant frequencies between 45 and 89 mutants per 1E+6) in a series of data points between 100% to 25% survival and there was also a negative data point between 10% and 1% survival (Office of Food Additive Safety, 2001). In this case it would be acceptable to count the TFT colonies of cultures exhibiting < 10% total growth. - Statistics:
- The cytotoxic effects of each treatment condition were expressed relative to the solvent-treated control for suspension growth over two days post-treatment and for total growth (suspension growth corrected for plating efficiency at the time of selection). The mutant frequency (number of mutants per 1E+6 surviving cells) for each treatment condition was determined by dividing the average number of colonies in the three TFT plates by the average number of colonies in the three corresponding VC plates and multiplying by the dilution factor (2 x 10-4) then multiplying by 1E+6. For simplicity, this was described as: (Average # TFT colonies / average # VC colonies) x 200 in the tables. The induced mutant frequency (IMF) was defined as the mutant frequency of the treated culture minus the mutant frequency of the solvent control cultures. The International Workshop on Genotoxicity established a Global Evaluation Factor (GEF) for a positive response at an IMF of ≥ 90 mutants per 10E+6 clonable cells at the Aberdeen meeting in 2003, published in Moore et al., 2006.
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- -S9 mix: ≥ 30 µg/mL; +S9: ≥ 50 µg/mL
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: in the preliminary experiment, visible precipitate was present in the treatment medium at concentrations ≥ 500 μg/mL at the beginning of treatment, at concentrations ≥ 1500 μg/mL at the end of the 4-hour treatment period and at a concentration of 5000 μg/mL at the end of the 24-hour treatment period.
RANGE-FINDING/SCREENING STUDIES: in the preliminary range-finding test, overt cytotoxicity (relative suspension growth ≤ 10%) was observed after 4 h at ≥ 50 µg/mL with and without metabolic activation. The suspension growth relative to the solvent control was 0% at concentrations ≥ 150 μg/mL in the presence and absence of S9 activation after the 4-hour treatment and at concentrations ≥ 50 μg/mL in the absence of S9 activation after the 24-hour treatment. Based on the results of the toxicity test, the test article concentrations selected for the initial mutagenesis assay with a 4-hour treatment ranged from 10 to 150 μg/mL for the non-activated and S9-activated cultures. The test article concentrations in the extended treatment assay with a 24-hour treatment ranged from 5 to 75 μg/mL for the non-activated cultures.
COMPARISON WITH HISTORICAL CONTROL DATA: mutant frequencies of the negative and positive controls were within their respective historical ranges.
ADDITIONAL INFORMATION ON CYTOTOXICITY: in the 4-h experiment, significant cytotoxicity was evident at ≥ 30 µg/mL in the absence of S9 mix and at 50 µg/mL in the presence of S9 mix. Exposure to the test substance for 24 h in the absence of S9 mix resulted in marked cytotoxicity at 40 µg/mL. Only little toxicity was noted at the other test concentrations with and without metabolic activation at the different exposure durations. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results:
negative - Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Remarks:
- Summary of available data used for the endpoint assesssment of the target substance
- Adequacy of study:
- key study
- Justification for type of information:
- refer to analogue justification provided in IUCLID section 13
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 5000 µg/plate: TA 1535 (+S9)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 5000 µg/plate: TA 1537 (-S9)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- As detailed in the analogue justification, it is considered that the target and the source substance are unlikely to lead to differences in in vitro bacterial mutagenicity potential. The result of the available AMES test with the source substance 3-(2,3-dihydroxypropoxy)-2-hydroxypropyl octadec-9-enoate was negative. Therefore, the target substance is also not expected to show mutagenic properties in bacteria.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Remarks:
- Summary of available data used for the endpoint assesssment of the target substance
- Adequacy of study:
- key study
- Justification for type of information:
- refer to analogue justification provided in IUCLID section 13
- Reason / purpose for cross-reference:
- read-across source
- Target gene:
- not applicable
- Key result
- Species / strain:
- other: human peripheral blood lymphocytes
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 150 μg/mL in the 4-hour nonactivated test condition, at 100 μg/mL in the 22-hour non-activated test and in the 4-hour S9-activated test at 200 µg/mL and above
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- As detailed in the analogue justification, it is considered that the target and the source substance are unlikely to lead to differences in in vitro mammalian chromosome aberration potential. The result of the available in vitro mammalian chromosome aberration test with the source substance 3-(2,3-dihydroxypropoxy)-2-hydroxypropyl octadec-9-enoate was negative. Therefore, the target substance is not expected to show clastogenic properties.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Remarks:
- Summary of available data used for the endpoint assesssment of the target substance
- Adequacy of study:
- key study
- Justification for type of information:
- refer to analogue justification provided in IUCLID section 13
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- -S9 mix: ≥ 30 µg/mL; +S9: ≥ 50 µg/mL
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- As detailed in the analogue justification, it is considered that the target and the source substance are unlikely to lead to differences in in vitro gene mutation in mammalian cells. The result of the available in vitro gene mutation in mammalian cells with the source substance 3-(2,3-dihydroxypropoxy)-2-hydroxypropyl octadec-9-enoate was negative was negative. Therefore, the target substance is also not expected to show mutagenic properties in mammalian cells.
Referenceopen allclose all
Table1. Test results of first experiment – preliminary cytotoxicity test
Bacterial Reverse Mutation Assay, mean revertant colonies/plate (n=2 ± SD) |
|||||
EXPERIMENT I (plate incorporation) |
|||||
S9-Mix
|
Without
|
||||
Concentration (per plate) |
TA 98 |
TA 100 |
TA 1535 |
TA 1537 |
WP2 uvrA |
SC (DMSO) |
17 ± 1 |
111 ± 4 |
16 ± 0 |
5 ± 1 |
31 ± 1 |
Test item |
|
|
|
|
|
33.3 µg |
20 ± 4 |
98 ± 6 |
20 ± 2 |
14 ± 1 |
38 ± 7 |
66.7 µg |
24 ± 6 |
110 ± 11 |
11 ± 3 |
10 ± 1 |
33 ± 8 |
100 µg |
23 ± 6 |
101 ± 16 |
14 ± 7 |
3 ± 3 |
33 ± 4 |
333 µg |
25 ± 5 |
91 ± 0 |
14 ± 4 |
5 ± 0 |
35 ± 4 |
667 µg |
21 ± 4 |
84 ± 18 |
18 ± 4 |
4 ± 1 |
39 ± 6 |
1000 µg |
28 ± 1 |
100 ± 6 |
9 ± 6 |
7 ± 0 |
38 ± 14 |
3333 µg |
22 ± 4P |
80 ± 4P |
9 ± 6P |
10 ± 7P |
39 ± 8 |
5000 µg |
23 ± 7P |
89 ± 1P |
18 ± 0P |
6 ± 6P |
26 ± 4P |
PC |
|
|
|
|
|
2-NF (1.0 µg) |
143 ± 0 |
- |
- |
- |
- |
SA (2.0 µg) |
- |
1295 ± 50 |
1115 ± 115 |
- |
- |
ICR-191 (2.0 µg) |
- |
- |
- |
1440 ± 53 |
- |
4-NQO (1.0 µg) |
- |
- |
- |
- |
540 ± 13 |
S9-Mix
|
With |
||||
Concentration (per plate) |
TA 98 |
TA 100 |
TA 1535 |
TA 1537 |
WP2 uvrA |
SC (acetone) |
33 ± 8 |
139 ± 1 |
12 ± 2 |
15 ± 4 |
56 ± 2 |
Test item |
|
|
|
|
|
33.3 µg |
41 ± 9 |
132 ± 4 |
17 ± 1 |
11 ± 1 |
61 ± 4 |
66.7 µg |
39 ± 6 |
135 ± 8 |
11 ± 3 |
10 ± 1 |
49 ± 5 |
100 µg |
27 ± 4 |
140 ± 11 |
11 ± 1 |
13 ± 0 |
54 ± 8 |
333 µg |
33 ± 4 |
116 ± 7 |
11 ± 1 |
12 ± 3 |
55 ± 8 |
667 µg |
32 ± 4 |
73 ± 7 |
12 ± 2 |
11 ± 4 |
45 ± 4 |
1000 µg |
34 ± 0 |
87 ± 20 |
11 ± 1 |
12 ± 6 |
49 ± 1 |
3333 µg |
31 ± 6P |
80 ± 9 |
10 ± 4P |
5 ± 6P |
47 ± 4 |
5000 µg |
25 ± 1P |
94 ± 16P |
8 ± 2P |
7 ± 1P |
51 ± 8 |
PC |
|
|
|
|
|
BP (2.5 µg) |
485 ± 87 |
- |
- |
- |
- |
2-AA (2.5 µg) |
- |
2501 ± 137 |
196 ± 1 |
181 ± 45 |
- |
2-AA (25 µg) |
- |
- |
- |
- |
257 ± 8 |
SC = Solvent control; PC = Positive control substances; SD = standard deviation; BaP: benzo[a]pyrene; 4-NQO: 4-nitroquinoline N-oxide; acridine mutagen ICR-191: ICR-191; SA: sodium azide; 2-aminoanthracene: 2-AA; 2-NF: 2-nitrofluorene P = precipitate; M = manual counting necessary |
|||||
Table 2. Test results of second experiment – Main assay
Bacterial Reverse Mutation Assay, mean revertant colonies/plate (n=3 ± SD) |
|||||
EXPERIMENT II (plate incorporation) |
|||||
S9-Mix
|
Without
|
||||
Concentration (per plate) |
TA 98 |
TA 100 |
TA 1535 |
TA 1537 |
WP2 uvrA |
SC (DMSO) |
18 ± 2 |
86 ± 7 |
11 ± 8 |
8 ± 2 |
25 ± 3 |
Test item |
|
|
|
|
|
333 µg |
18 ± 3 |
74 ± 2 |
6 ± 4 |
5 ± 1 |
25 ± 5 |
667 µg |
14 ± 3 |
74 ± 3 |
9 ± 3 |
4 ± 1 |
28 ± 6 |
1000 µg |
16 ± 1 |
69 ± 2 |
8 ± 3 |
5 ± 1 |
24 ± 2 |
3333 µg |
18 ± 5P |
78 ± 16P |
9 ± 3P |
6 ± 3P |
26 ± 4 |
5000 µg |
19 ± 3P |
74 ± 3P |
6 ± 4P |
3 ± 1P, M |
26 ± 3P |
PC |
|
|
|
|
|
2-NF (1.0 µg) |
150 ± 3 |
- |
- |
- |
- |
SA (2.0 µg) |
- |
1031 ± 19 |
956 ± 37 |
- |
- |
ICR-191 (2.0 µg) |
- |
- |
- |
1649 ± 123 |
- |
4-NQO (1.0 µg) |
- |
- |
- |
- |
456 ± 90 |
S9-Mix
|
With |
||||
Concentration (per plate) |
TA 98 |
TA 100 |
TA 1535 |
TA 1537 |
WP2 uvrA |
SC (acetone) |
28 ± 6 |
99 ± 11 |
15 ± 2 |
5 ± 1 |
33 ± 1 |
Test item |
|
|
|
|
|
333 µg |
27 ± 1 |
94 ± 24 |
8 ± 2 |
9 ± 2 |
32 ± 5 |
667 µg |
23 ± 4 |
72 ± 5 |
7 ± 3 |
6 ± 1 |
29 ± 3 |
1000 µg |
20 ± 4P |
72 ± 13 |
8 ± 2 |
5 ± 4 |
29 ± 4 |
3333 µg |
19 ± 2P |
63 ± 6P |
8 ± 3P |
5 ± 3P |
35 ± 9P |
5000 µg |
22 ± 6P |
63 ± 7P |
6 ± 4P |
4 ± 2P |
30 ± 10P |
PC |
|
|
|
|
|
BP (2.5 µg) |
284 ± 12 |
- |
- |
- |
- |
2-AA (2.5 µg) |
- |
2337 ± 174 |
155 ± 5 |
119 ± 22 |
- |
2-AA (25 µg) |
- |
- |
- |
- |
200 ± 19 |
SC = Solvent control; PC = Positive control substances; SD = standard deviation; BaP: benzo[a]pyrene; 4-NQO: 4-nitroquinoline N-oxide; acridine mutagen ICR-191: ICR-191; SA: sodium azide; 2-aminoanthracene: 2-AA; 2-NF: 2-nitrofluorene P = precipitate |
|||||
Table 1: Cytogenic analysis of cells treated with the test substance in the presence or absence of exogenous metabolic acitvation
|
Cell scored |
Aberrations per Cell |
Cells with Aberrations# |
||||||
Treatment (µg/mL) |
S9 activation |
Treatment time |
Mitotic index (%) |
Numerical |
Structural |
Mean |
SD |
Numerical (%) |
Structural (%) |
Vehicle |
- |
4 |
14.0 |
200 |
200 |
0.005 |
0.007 |
0.0 |
0.5 |
25 |
- |
4 |
12.3 |
200 |
200 |
0.000 |
0.000 |
0.0 |
0.0 |
50 |
- |
4 |
11.7 |
200 |
200 |
0.000 |
0.000 |
0.0 |
0.0 |
100 |
- |
4 |
8.6 |
200 |
200 |
0.030 |
0.028 |
1.0 |
2.0 |
150 |
- |
4 |
3.6 |
200 |
200 |
0.015 |
0.007 |
0.0 |
1.5 |
MMC 0.2 |
- |
4 |
7.6 |
200 |
200 |
0.080 |
0.042 |
0.0 |
8.0* |
Vehicle |
+ |
4 |
11.6 |
200 |
200 |
0.010 |
0.014 |
0.0 |
1.0 |
50 |
+ |
4 |
11.0 |
200 |
200 |
0.005 |
0.007 |
1.5 |
0.5 |
100 |
+ |
4 |
10.0 |
200 |
200 |
0.010 |
0.014 |
0.0 |
1.0 |
200 |
+ |
4 |
6.3 |
200 |
200 |
0.020 |
0.014 |
0.0 |
2.0 |
CP 10 |
+ |
4 |
7.0 |
200 |
200 |
0.125 |
0.035 |
0.0 |
10.5* |
Vehicle |
- |
22 |
8.0 |
200 |
200 |
0.025 |
0.007 |
0.0 |
2.5 |
25 |
- |
22 |
6.5 |
200 |
200 |
0.000 |
0.000 |
0.0 |
0.0 |
50 |
- |
22 |
5.2 |
200 |
200 |
0.005 |
0.007 |
0.0 |
0.5 |
100 |
- |
22 |
2.6 |
200 |
200 |
0.010 |
0.014 |
0.0 |
1.0 |
MMC 0.2 |
- |
22 |
5.1 |
200 |
200 |
0.135 |
0.007 |
0.0 |
12.5* |
*: Statistically significant difference from control at p < 0.05 by Fisher's test.
#: Excluding cells with only gaps
Table 1. Experiment I - 4 h exposure - With Metabolic Activation
Concentration [µg/mL] |
Suspension growth [%] |
Relative total growth [%] |
Mutant frequency per 1E+06 surviving cells |
IMF per 1E+06 surviving cells* |
|
SC |
100 |
100 |
42 |
N/A |
|
SC |
59 |
||||
10 |
107 |
95 |
61 |
11 |
|
10 |
101 |
93 |
50 |
0 |
|
20 |
106 |
109 |
40 |
-10 |
|
20 |
102 |
94 |
55 |
4 |
|
30 |
107 |
96 |
59 |
9 |
|
30 |
95 |
99 |
44 |
-6 |
|
40 |
79 |
78 |
58 |
7 |
|
40 |
70 |
76 |
57 |
7 |
|
50 |
19 |
18 |
65 |
14 |
|
50 |
23 |
18 |
71 |
21 |
|
DMBA, 1.5 |
13 |
7 |
560 |
510 |
|
DMBA, 1.25 |
12 |
7 |
532 |
482 |
DMBA = 7,12-dimethylbenzanthracene; SC = solvent control (DMSO); IMF = Induced mutant frequency
* compared to a mean solvent mutant frequency of 50 per 10E+06 cells
Table 2. Experiment I - 3 h exposure - Without Metabolic Activation
Concentration [µg/mL] |
Suspension growth [%] |
Relative total growth [%] |
Mutant frequency per 1E+06 surviving cells |
IMF per 1E+06 surviving cells* |
|
SC |
100 |
100 |
65 |
N/A |
|
SC |
50 |
||||
10 |
103 |
89 |
72 |
14 |
|
10 |
104 |
93 |
49 |
-9 |
|
20 |
103 |
93 |
53 |
-5 |
|
20 |
96 |
88 |
44 |
-14 |
|
30 |
86 |
78 |
47 |
-11 |
|
30 |
57 |
48 |
57 |
-1 |
|
40 |
55 |
54 |
58 |
0 |
|
40 |
58 |
51 |
50 |
-8 |
|
50 |
20 |
18 |
45 |
-13 |
|
50 |
16 |
14 |
68 |
10 |
|
MMS, 20 |
43 |
7 |
1179 |
1122 |
|
MMS, 15 |
53 |
20 |
669 |
611 |
MMS = methylmethanesulfonate; SC = solvent control (DMSO); IMF = Induced mutant frequency
* compared to a mean solvent mutant frequency of 58 per 10E+06 cells
Table 3. Experiment II - 24 h exposure - Without Metabolic Activation
Concentration [µg/mL] |
Suspension growth [%] |
Relative total growth [%] |
Mutant frequency per 1E+06 surviving cells |
IMF per 1E+06 surviving cells* |
|
SC |
100 |
100 |
43 |
N/A |
|
SC |
47 |
||||
5 |
107 |
83 |
31 |
-4 |
|
5 |
100 |
72 |
31 |
-2 |
|
10 |
104 |
77 |
45 |
15 |
|
10 |
101 |
81 |
39 |
4 |
|
20 |
103 |
85 |
40 |
3 |
|
20 |
98 |
82 |
39 |
1 |
|
30 |
97 |
83 |
34 |
-5 |
|
30 |
97 |
79 |
38 |
1 |
|
40 |
30 |
27 |
49 |
9 |
|
40 |
33 |
28 |
59 |
21 |
|
MMS, 7.5 |
42 |
14 |
268 |
696 |
|
MMS, 5 |
52 |
27 |
256 |
394 |
MMS = methylmethanesulfonate; SC = solvent control (DMSO); IMF = Induced mutant frequency
* compared to a mean solvent mutant frequency of 40 per 10E+06 cells
RESULTS OF COLONY SIZING
The size of the colonies was not determined for test substance-treated cultures, since no positive results for mutagenicity were obtained. The colony sizing for the MMS and DMBA positive controls yielded the expected increase in small colonies (verifying the adequacy of the methods used to detect small colony mutants) and large colonies.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
There are no genetic toxicity data available on Fatty acids, C16-18 (even numbered), esters with glycerol oligomers. However, in EFSA’s scientific opinion on the re-evaluation of polyglycerol esters of fatty acids (PEPA, E 475), as a food additive, it is stated that the safety of polyglycerols and specific fatty acids has recently been assessed and no adverse effects were identified in the available studies. No genotoxic potential of PEFA was identified from the limited information available (EFSA, 2017). Genetic toxicity studies with the registered substance have been commissioned at a CRO. Therefore, as an interim measure, the genetic toxicity potential of the registered substance was assessed based on the available data from the source substance oleic acid, monoester with oxybis(propanediol), (CAS 49553-76-6). In accordance with Regulation (EC) No. 1907/2006 Annex XI, 1.5 “Grouping of substances and read across” and following the Read across assessment framework (RAAF, ECHA 2017) read across from an analogue substance has been applied to support the human health hazard assessment of Fatty acids, C16-18 (even numbered), esters with glycerol oligomers.
CAS 49553-76-6
Genetic toxicity (mutagenicity) in bacteria in-vitro
A bacterial gene mutation assay (Ames test) was performed with oleic acid, monoester with oxybis(propanediol), (CAS 49553-76-6) according to OECD Guideline 471 and under GLP conditions (Myhre, 2012). The plate incorporation procedure was performed with Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 and E. coli WP2 uver A strain in the absence and presence of metabolic activation (Aroclor 1254-induced rat liver S9-mix). A preliminary cytotoxicity test and a main assay were conducted each in triplicates at concentrations from 33.3 to 5000 µg/plate and 333 to 5000 µg/plate respectively (vehicle: DMSO). 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. Under the study conditions, the test substance did not induce mutations in the bacterial mutation assay in the absence and presence of a metabolic activation system in any of the strains tested.
Genetic toxicity (mutagenicity) in mammalian cells in-vitro
The in vitro mammalian cell gene mutation study of with oleic acid, monoester with oxybis(propanediol), (CAS 49553-76-6) was carried out according to OECD Guideline 476 under GLP conditions (Clarke, 2012). Gene mutations in the thymidine kinase locus were investigated in L5178Y mouse lymphoma cells in the presence and absence of a metabolic activation system (Aroclor 1254 rat liver S9). In the preliminary toxicity assay, the maximum concentration of test substance in medium was 5000 µg/mL. In the first experiment, cells were exposed for 4 h to test substance at concentrations of 10-150 µg/mL (in DMSO) with and without metabolic activation. The concentrations chosen for cloning were 10, 20, 30, 40 and 50 µg/mL in the presence and absence of metabolic activation. No cloned cultures exhibited induced mutant frequencies ≥ 90 mutants per 10 E6 clonable cells.
Concentrations of the second experiment without metabolic activation for an exposure time of 24 h ranged from 5-75 µg/mL. The concentrations chosen for cloning were 5, 10, 20, 30 and 40 µg/mL. No cloned cultures exhibited induced mutant frequencies ≥ 90 mutants per 10E6 clonable cells. The vehicle and positive controls in the study showed the expected results and were within the range of historical control data. Cytotoxicity was observed at concentrations ≥ 30 µg/mL without metabolic activation and ≥ 50 µg/mL with metabolic activation. 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, oleic acid, monoester with oxybis(propanediol) did not show gene mutation activity in this test performed in L5178Y mouse lymphoma cells in vitro.
Genetic toxicity (cytogenicity) in mammalian cells in-vitro
CAS 49553-76-6
An in vitro mammalian chromosome aberration test was performed with oleic acid, monoester with oxybis(propanediol), in cultured peripheral human lymphocytes comparable to OECD Guideline 473 and under GLP conditions (Glover, 2012). Duplicate cultures of human lymphocytes were evaluated for chromosome aberrations in the presence and absence of metabolic activation (rat liver S9-mix). Cells were exposed for 4 hours to the test substance dissolved in DMSO at concentrations of 25, 50, 100, 150, 200 µg/mL without metabolic activation, for 4 hours at concentration of 50, 100, 200, 300 and 400 µg/mL with metabolic activation and for 22 hours without metabolic activation at concentration of 10, 25, 50, 75 and 100 µg/mL. The test substance induced cytotoxicity at 150 μg/mL in the 4-hour non activated test condition, at 100 μg/mL in the 22-hour non-activated test and in the 4-hour S9-activated test at 200 µg/mL and above. Vehicle (solvent) controls induced aberration frequencies within the range expected for normal human lymphocytes. Mitomycin C and Cyclophosphamide were used as positive control substances inducing statistically significant increases in aberration frequencies indicating the satisfactory performance of the test and of the activity of the metabolizing system. Evaluation of 100 well-spread metaphase cells from each culture for structural chromosomal aberrations revealed no increase in the frequency of chromosome aberrations and polyploid cells at any dose level tested in comparison to the negative controls. The test material was therefore considered to be non-clastogenic to human lymphocytes in vitro.
Conclusions
In summary, one study investigating the genetic mutation in bacteria in-vitro is available with the source substance oleic acid, monoester with oxybis(propanediol) (CAS 49553-76-6) providing negative results. Furthermore, no cytogenicity in mammalian cells in-vitro and no mutagenicity in mammalian cells in-vitro were observed with the source substance oleic acid, monoester with oxybis(propanediol), (CAS 49553-76-6). Thus and based on read across, it can be concluded that the registration substance does not bear ant mutagenic and/or genotoxic properties.
Justification for classification or non-classification
Based on read-across from structurally similar substances, the available data on mutagenicity and genotoxicity do not meet the classification criteria according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, 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.
