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EC number: 500-129-7 | CAS number: 55552-95-9
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
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- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
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- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
All criteria for a valid study were met as described in the protocol. The results of the Bacterial Reverse Mutation Assay indicate that, under the conditions of this study, EK111 did not cause a positive mutagenic response with any of the tester strains in either the presence or absence of Aroclor-induced rat liver S9. The study was concluded to be negative without conducting a confirmatory (independent repeat) assay because the results were clearly negative; hence, no further testing was warranted.
Under the conditions of the assay described in this report, EK111 was concluded to be positive for the induction of forward mutations at the thymidine kinase locus in L5178Y mouse lymphoma cells, in the 4- hour treatment condition in the absence of an exogenous metabolic activation system, in the in vitro L5178Y/TK+/- mouse lymphoma assay.
Under the conditions of the assay described in this report, EK111 was concluded to be negative for the induction of structural and numerical chromosomal aberrations in the non-activated and S9-activated test systems in the in vitro mammalian chromosomal aberration test using CHO cells.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Vehicle / solvent:
- DMSO
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- methylmethanesulfonate
- other: 2-aminoanthracene
- Evaluation criteria:
- All Salmonella tester strain cultures must demonstrate the presence of the deep rough mutation (rfa) and the deletion in the uvrB gene. Cultures of tester strains TA98 and TA100 must demonstrate the presence of the pKM101 plasmid R-factor. All WP2 uvrA cultures must demonstrate the deletion in the uvrA gene.
All cultures must demonstrate the characteristic mean number of spontaneous revertants in the vehicle controls as follows (inclusive): TA98, 10 - 50; TA100, 80 - 240; TA1535, 5 - 45; TA1537, 3 - 21; WP2 uvrA, 10 - 60.
To ensure that appropriate numbers of bacteria are plated, tester strain culture titers must be greater than or equal to 0.3x109 cells/mL.
The mean of each positive control must exhibit at least a 3.0-fold increase in the number of revertants over the mean value of the respective vehicle control.
A minimum of three non-toxic dose levels is required to evaluate assay data. A dose level is considered toxic if one or both of the following criteria are met: (1) A >50 % reduction in the mean number of revertants per plate as compared to the mean vehicle control value. This reduction must be accompanied by an abrupt dose-dependent drop in the revertant count. (2) At least a moderate reduction in the background lawn (background code 3, 4 or 5). - 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 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:
- E. coli WP2 uvr A
- Metabolic activation:
- with
- 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
- Conclusions:
- All criteria for a valid study were met as described in the protocol. The results of the Bacterial Reverse Mutation Assay indicate that, under the conditions of this study, EK111 did not cause a positive mutagenic response with any of the tester strains in either the presence or absence of Aroclor-induced rat liver S9. The study was concluded to be negative without conducting a confirmatory (independent repeat) assay because the results were clearly negative; hence, no further testing was warranted.
- Executive summary:
The test substance, EK111, was tested to evaluate its mutagenic potential by measuring its ability to induce reverse mutations at selected loci of several strains of Salmonella typhimurium and at the tryptophan locus of Escherichia coli strain WP2 uvrA in the presence and absence of an exogenous metabolic activation system. Dimethyl sulfoxide (DMSO) was used as the vehicle.
In the preliminary toxicity assay, the dose levels tested were 6.67, 10.0, 33.3, 66.7, 100, 333, 667, 1000, 3333 and 5000 μg per plate. No precipitate was observed. Toxicity was observed beginning at concentrations from 333 to 3333 μg per plate. Based upon these results, the maximum dose tested in the mutagenicity assay was 3333 μg per plate with all Salmonella tester strains in the absence of S9 activation and tester strains TA100 and TA1535 in the presence of S9 activation and 5000 μg per plate with the remaining test conditions.
In the mutagenicity assay, the dose levels tested were 10.0, 33.3, 100, 333, 1000 and 3333 μg per plate with all Salmonella tester strains in the absence of S9 activation and tester strains TA100 and TA1535 in the presence of S9 activation and 33.3, 100, 333, 1000, 3333 and 5000 μg per plate with the remaining test conditions. No precipitate was observed. Toxicity was observed beginning at 1000 or 3333 μg per plate. No positive mutagenic responses were observed with any of the tester strains in either the presence or absence of S9 activation.
These results indicate EK111 was negative for the ability to induce reverse mutations at selected loci of several strains of Salmonella typhimurium and at the tryptophan locus of Escherichia coli strain WP2 uvrA in the presence and absence of an exogenous metabolic activation system.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 490 (In Vitro Mammalian Cell Gene Mutation Tests Using the Thymidine Kinase Gene)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian cell transformation assay
- Target gene:
- thymidine kinase
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254-induced rat liver S9
- Test concentrations with justification for top dose:
- Non-activated 4 hours 9.4, 18.8, 37.5, 75, 150, 300 and 600
Non-activated 24 hours 1, 2, 4, 8, 15, 20 and 40
S9-activated 4 hours 18.8, 37.5, 75, 150, 300, 600 and 800
Based upon the results of the preliminary toxicity assay - Vehicle / solvent:
- DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- methylmethanesulfonate
- Details on test system and experimental conditions:
- The preparation and addition of the test substance dose formulations was carried out under filtered lighting during the exposure period. Treatment was carried out by combining 100 μL of test substance dose formulation, vehicle or positive control dose formulation and F0P medium or S9 mix (as appropriate) with 6 x 10^6 L5178Y/TK+/- cells in a total volume of 10 mL. All pH adjustments were performed prior to adding S9 or target cells to the treatment medium. Each S9-activated 10-mL culture contained 4 mL S9 mix (final S9 concentration of 1.0%). Cultures were capped tightly and incubated with mechanical mixing at 37 ± 1°C for 4 or 24 hours.
For the preliminary toxicity assay only, after a 4-hour treatment in the presence and absence of S9, cells were washed with culture medium and cultured in suspension for two days post-treatment, with cell concentration adjustment on the first day. After a 24-hour treatment in the absence of S9, cells were washed with culture medium and immediately readjusted to 3 x 105 cells/mL. Cells were then cultured in suspension for an additional two days post-treatment with cell concentration adjustment on the first day.
For the definitive assay only, at the end of the exposure period, the cells were washed with culture medium and collected by centrifugation. The cells were resuspended in 20 mL F10P on Day 1 and in 10 mL F10P on Day 2, and incubated at 37 ± 1°C for two days following treatment. Cell population adjustments to 3 x 10^5 cells/mL were made as follows:
• 4 hour treatment – 1 and 2 days after treatment.
• 24 hour treatment – immediately after test substance removal, and 2 and 3 days after treatment. - Evaluation criteria:
- • A result was considered positive if a concentration-related increase in 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/10^6 clonable cells (based on the average mutant frequency of duplicate cultures). If the average vehicle control mutant frequency was >90 mutants/10^6 clonable cells, a doubling of mutant frequency over the vehicle would 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/10^6 clonable cells (based on the average mutant frequency of duplicate cultures) and there was no concentration-related increase in mutant frequency. - Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Under the conditions of the assay described in this report, EK111 was concluded to be positive for the induction of forward mutations at the thymidine kinase locus in L5178Y mouse lymphoma cells, in the 4- hour treatment condition in the absence of an exogenous metabolic activation system, in the in vitro L5178Y/TK+/- mouse lymphoma assay.
- Executive summary:
The test substance, EK111, was evaluated for its ability to induce forward mutations at the thymidine kinase locus in L5178Y mouse lymphoma cells in the presence and absence of an exogenous metabolic activation system. Dimethyl sulfoxide (DMSO) was used as the vehicle.
In the preliminary toxicity assay, the concentrations tested were 19.5, 39.1, 78.1, 156, 313, 625, 1250, 2500 and 5000 μg/mL. The maximum concentration evaluated approximated the limit dose for this assay. No visible precipitate was observed at the beginning or end of treatment. Relative suspension growth (RSG) was 11, 12 and 13% at concentrations of 625 μg/mL (4-hour treatment with S9), 313 μg/mL (4-hour treatment without S9) and 19.5 μg/mL (24-hour treatment without S9), respectively. RSG was 0% at all higher concentrations using all treatment conditions. Based upon these results, the concentrations chosen for the definitive mutagenicity assay were 18.8, 37.5, 75, 150, 300, 600 and 800 μg/mL (4-hour treatment with S9), 9.4, 18.8, 37.5, 75, 150, 300 and 600 μg/mL (4-hour treatment without S9) and 1, 2, 4, 8, 15, 20 and 40 μg/mL (24-hour treatment without S9).
In the initial definitive mutagenicity assay, no visible precipitate was observed at the beginning or end of treatment. Cultures treated at concentrations of 18.8, 37.5, 75, 300 and 600 μg/mL (4-hour treatment with S9) and 1, 2, 4, 8 and 15 μg/mL (24 -hour treatment without S9) exhibited 24 to 112% and 16 to 86% RSG, respectively, and were cloned. Relative total growth of the cloned cultures ranged from 18 to 125% (4-hour treatment with S9) and 13 to 95% (24-hour treatment without S9). Cultures treated at other concentrations were discarded prior to cloning because a sufficient number of other concentrations were available or due to excessive toxicity. No increases in induced mutant frequency ≥90 mutants/106 clonable cells were observed under any treatment condition. The 4-hour treatment without S9 was discontinued as the Solvent Colony Efficiency was outside the acceptable range. Based upon these results, the concentrations chosen for the retest of definitive mutagenicity assay for 4-hour treatment without S9 were the same as initial definitive mutagenicity assay.
In the retest of definitive mutagenicity assay for 4-hour treatment without S9, no visible precipitate was observed at the beginning or end of treatment. Cultures treated at concentrations of 9.4, 18.8, 37.5, 75 and 150 μg/mL exhibited 13 to 99 %RSG, and were cloned. Relative total growth of the cloned cultures ranged from 10 to 96 % (One of the cultures at 150 μg/mL was excluded from evaluation of mutagenicity due to excessive toxicity). Cultures treated at other concentrations were discarded prior to cloning due to excessive toxicity. Increases in induced mutant frequency ≥90 mutants/106 clonable cells were observed at concentrations ≥75 μg/mL.
These results indicate EK111 was positive for 4-hour treatment without S9 for the ability to induce forward mutations at the thymidine kinase locus in L5178Y mouse lymphoma cells, in the presence and absence of an exogenous metabolic activation system.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian cell micronucleus test
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor 1254-induced rat liver S9
- Test concentrations with justification for top dose:
- Treatment Condition Treatment Time Recovery Time Doses (μg/mL)
Non-activated 4 hr 16 hr 5, 10, 20, 30, 35, 40, 45, 50, 60
20 hr 0 hr 5, 10, 20, 30, 35, 40, 45, 50, 60
S9-activated 4 hr 16 hr 30, 50, 100, 125, 150, 200, 300, 450, 500 - Vehicle / solvent:
- DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- methylmethanesulfonate
- Details on test system and experimental conditions:
- Chinese hamster ovary (CHO-K1) cells (repository number CCL 61) were obtained from American Type Culture Collection, Manassas, VA. In order to assure the karyotypic stability of the cell line, working cell stocks were not used beyond passage 15. The frozen lot of cells was tested using the Hoechst staining procedure and found to be free of mycoplasma contamination. This cell line has an average cell cycle time of 10-14 hours with a modal chromosome number of 20. The use of CHO cells has been demonstrated to be an effective method of detection of chemical clastogens (Preston et al., 1981).
- Evaluation criteria:
- The test substance was considered to have induced a positive response if
• at least one of the test concentrations exhibits a statistically significant increase when compared with the concurrent negative control (p ≤ 0.05), and
• the increase is concentration-related (p ≤ 0.05), and
• results are outside the 95% control limit of the historical negative control data.
The test substance was considered to have induced a clear negative response if none of the criteria for a positive response were met. - Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Under the conditions of the assay described in this report, EK111 was concluded to be negative for the induction of structural and numerical chromosomal aberrations in the non-activated and S9-activated test systems in the in vitro mammalian chromosomal aberration test using CHO cells.
- Executive summary:
The test substance, EK111, was tested to evaluate the potential to induce structural chromosomal aberrations using Chinese hamster ovary (CHO) cells in both the absence and presence of an of an exogenous metabolic activation system. CHO cells were treated for 4 hours in the absence and presence of S9, and for 20 hours in the absence of S9. Dimethyl sulfoxide (DMSO) was used as the vehicle.
In the preliminary toxicity assay, the doses tested ranged from 0.5 to 5000 μg/mL, which was the limit dose for this assay. Cytotoxicity (≥ 50% reduction in cell growth index relative to the vehicle control) was observed at doses ≥ 50 μg/mL in the non-activated 4 and 20-hour exposure groups; and at doses ≥ 150 μg/mL in the S9-activated 4-hour exposure group. At the conclusion of the treatment period, visible precipitate was observed at doses ≥ 500 μg/mL in the S9-activated 4-hour exposure group. Based upon these results, the doses chosen for the chromosomal aberration assay ranged from 5 to 60 μg/mL for the non-activated 4 and 20-hour exposure groups; and from 30 to 500 μg/mL for the S9-activated 4-hour exposure group.
In the chromosomal aberration assay, cytotoxicity (≥ 50% reduction in cell growth index relative to the vehicle control), was observed at doses ≥ 45 μg/mL in the non-activated 4-hour exposure group; at doses ≥ 50 μg/mL in the S9-activated 4-hour exposure group; and at doses ≥ 20 μg/mL in the non-activated 20-hour exposure group. The doses selected for evaluation of chromosomal aberrations were 10, 20, and 45 μg/mL for the non-activated 4-hour exposure group; 30, 50, and 100 μg/mL for the S9-activated 4-hour exposure group; and 5, 10, and 20 μg/mL for the non-activated 20-hour exposure group.
No significant or dose-dependent increases in structural or numerical (polyploid or endoreduplicated cells) aberrations were observed in treatment groups with or without S9 (p > 0.05; Fisher’s Exact and Cochran-Armitage tests).
These results indicate EK111 was negative for the induction of structural and numerical chromosomal aberrations in the presence and absence of the exogenous metabolic activation system.
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