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EC number: 500-038-2 | CAS number: 25322-68-3 1 - 4.5 moles ethoxylated
- 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
- Surface tension
- 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
- Biodegradation
- Bioaccumulation
- Transport and distribution
- 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
Ames test:
Gene mutation toxicity study was performed for the test chemical to evaluate its mutagenic nature. The study was performed as per the preincubation protocol using Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system at doses of 0, 100, 333, 1000, 3333 or 10000 µg/plate. Water was used at the vehicle. The plates were incubated for 48 hrs after 20 mins preincubation before the evaluation of the revertant colonies could be made. Concurrent solvent and negative control chemicals were also included in the study. The test chemical did notinduce mutation in theSalmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.
In vitro mammalian chromosome aberration study:
In vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical. The study was performed using CHEL cellsin the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level of 0, 2, 3, 4.5, 6 or 8 mM. Concurrent solvent, untreated and positive control chemicals were also included in the study. 80000 CHEL cells were seeded in 25 cm2 flasks 9-20 hrs before treatment under standard culture conditions. The test chemical treatment was performed continuously for 16 or 24 hrs until harvesting. During the last 2 hrs of culture, colcemid was added. Cells were then collected by trypsin/EDTA treatment. Treated with hypotonic solution of tri-sodium citrate and washed twice with fresh fixative, dropped onto glass slides and air dried. Slides were stained with aqueous solution of 1% Giemsa and coded for analysis. Abpout 100-200 well spread metaphase cells were scored per test point. Based on the observations made, the test chemical did not induce chromosome aberrations in CHEL cell line at dose level of 2 mM. It however induced gene mutation in CHEL cells in the presence and absence of S9 metabolic activation system above 2 mM.
In vitro mammalian cell gene mutation assay:
An in vitro mammalian cell gene mutation study was designed and conducted to determine the genotoxicity profile of the test chemical when administered to Chinese Hamster Ovary (CHO) cells. In the genotoxicity test, the test chemical was administered to CHO cells for 3 hrs at the dose levels of 0.5, 1.0, 2.5 or 5.0 mM and in the absence or presence of exogenous metabolic activation. CHO cells representing the negative controls were exposed to the vehicle. Positive controls, such asN-ethyl-N-nitrosourea (ENU) experiments without metabolic activation and 7,12-dimethylbenz(a) anthracene in experiments with metabolic activation, were also included in each test. The positive control ENU showed indication of gene mutations occurring while no other treatment gave rise to gene toxicity. When the mutation frequency was determined, a frequency of 3.79 x 10-4was shown after a 3 hour exposure of ENU as the positive control and in the absence of S9 liver microsomal fraction. Since no tested concentration of Polyethylene glycol, in the absence or presence of S9 liver microsomal fraction, resulted in colonies, we conclude that Polyethylene glycol does not give rise to gene mutations when CHO cells are exposedin vitroto the test chemical at 0, 0.5, 1.0, 2.5 or 5.0 mM for 3 hrs.Based on the results of the current study, it is concluded that the test chemical does not give rise to gene mutations when CHO cells are exposed to the test chemicalin vitroat 0, 0.5, 1.0, 2.5 or 5.0 mM for 3 hrs, in the presence or abscence of metabolic activation.
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
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Justification for type of information:
- Data is from peer reviewed publication
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Principles of method if other than guideline:
- Gene mutation toxicity study was performed for the test chemical to evaluate its mutagenic nature
- GLP compliance:
- not specified
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- with and without
- Metabolic activation system:
- Male Sprague-Dawley rats and male Syrian hamsters were routinely used for the S9 preparation of the liver fractions
- Test concentrations with justification for top dose:
- 0, 100, 333, 1000, 3333 or 10000 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Water
- Justification for choice of solvent/vehicle: The test chemical was soluble in water - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Water
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- For strains tested with S9
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Water
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- For strains TA100 and TA1535 tested in the absence of S9
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Water
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- For strain TA1537 tested in the absence of S9
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Water
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- Remarks:
- For strain TA98 tested in the absence of S9
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: preincubation
DURATION
- Preincubation period: 20 mins
- Exposure duration: 48 hr
- Expression time (cells in growth medium): 48 hr
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data
SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data
NUMBER OF REPLICATIONS: At least five dose levels of the chemicals were tested, with three plates per dose level.
NUMBER OF CELLS EVALUATED: No data
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data
OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data
OTHER: No data - Rationale for test conditions:
- No data
- Evaluation criteria:
- 1) mutagenic response: a dose-related, reproducible increase in the number of revertants over background, even if the increase was less than twofold;
2) nomutagenic response: when no increase in the number of revertants was elicited by the chemical;
3) questionable response: when there was an absence of a clear-cut dose-related increase in revertants; when the dose-related increases in the number of revertants were not reproducible; or when the response was of insufficient magnitude to support a determination of mutagenicity - Statistics:
- Mean and Standard error of mean
- Species / strain:
- S. typhimurium, other: TA100, TA98, TA1535 and TA1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: No data
- Other confounding effects: No data
RANGE-FINDING/SCREENING STUDIES: The chemical was initially tested with strain TA100 in the presence and the absence of the metabolic activation systems, over a wide dose range with an upper limit of 10 mg/plate, or less when solubility problems were encountered. Toxicity was evidenced by one or more of the following phenomena: appearance of his+ pinpoint colonies, reduced numbers of revertant colonies per plate, or thinning or absence of the bacterial lawn. Nontoxic chemicals were tested in the initial experiment up to the 10 mg/plate dose level, or to a level determined by their solubility. Toxic chemicals were tested up to a high dose which exhibited some degree of toxicity.
COMPARISON WITH HISTORICAL CONTROL DATA: No data
ADDITIONAL INFORMATION ON CYTOTOXICITY: No data - Remarks on result:
- other: No mutagenic potential
- Conclusions:
- The test chemical did not induce mutation in the Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.
- Executive summary:
Gene mutation toxicity study was performed for the test chemical to evaluate its mutagenic nature. The study was performed as per the preincubation protocol using Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system at doses of 0, 100, 333, 1000, 3333 or 10000 µg/plate. Water was used at the vehicle. The plates were incubated for 48 hrs after 20 mins preincubation before the evaluation of the revertant colonies could be made. Concurrent solvent and negative control chemicals were also included in the study. The test chemical did notinduce mutation in theSalmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Justification for type of information:
- Data is from peer reviewed publication
- Qualifier:
- according to guideline
- Guideline:
- other: Refer below principle
- Principles of method if other than guideline:
- In vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical
- GLP compliance:
- not specified
- Type of assay:
- other: In vitro mammalian chromosome aberration study
- Target gene:
- No data
- Species / strain / cell type:
- mammalian cell line, other: CHEL cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: William’s medium supplemented with 10% fetal calf serum and antibiotics
- Properly maintained: No data
- Periodically checked for Mycoplasma contamination: No data
- Periodically checked for karyotype stability: The cell line has a stable aneuploidy karyotype with modal no. of chromosome of 24 and 21
- Periodically "cleansed" against high spontaneous background: No data - Additional strain / cell type characteristics:
- not specified
- Cytokinesis block (if used):
- No data
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 fraction was isolated from young male rats following mixed with β-naphthoflavone and phenobarbital
- Test concentrations with justification for top dose:
- 0, 2, 3, 4.5, 6 or 8 mM
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO - Untreated negative controls:
- yes
- Remarks:
- Untreated cells
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
Cell at the start of experiment: 80000
DURATION
- Preincubation period:
- Exposure duration: Continuous treatment: 16 hrs
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):
SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS:
NUMBER OF CELLS EVALUATED: 100-200 well spread metaphase per test point. Only metaphases with chromosome numbers that differed from the modal numbers by no more than two were considered.
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: Yes, cytotoxicity was assessed as a reduction in mitotic indices and the confluence status of the cultures
OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data
OTHER: No data - Rationale for test conditions:
- No data
- Evaluation criteria:
- The cell line was observed for chromosome breakages, gaps and exchanges
- Statistics:
- Statistical analysis of data was performed with Fischer’s exact test, accepting P<0.05 as significant
- Species / strain:
- mammalian cell line, other: CHEL cells
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Remarks:
- above 2 mM
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 13.3% at 8mM
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- mammalian cell line, other: CHEL cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- at 2 mM
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- No data
- Remarks on result:
- other: No mutagenic potential
- Conclusions:
- The test chemical did not induce chromosome aberrations in CHEL cell line at dose level of 2 mM. It however induced gene mutation in CHEL cells in the presence and absence of S9 metabolic activation system above 2 mM.
- Executive summary:
In vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical. The study was performed using CHEL cellsin the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level of 0, 2, 3, 4.5, 6 or 8 mM. Concurrent solvent, untreated and positive control chemicals were also included in the study. 80000 CHEL cells were seeded in 25 cm2 flasks 9-20 hrs before treatment under standard culture conditions. The test chemical treatment was performed continuously for 16 or 24 hrs until harvesting. During the last 2 hrs of culture, colcemid was added. Cells were then collected by trypsin/EDTA treatment. Treated with hypotonic solution of tri-sodium citrate and washed twice with fresh fixative, dropped onto glass slides and air dried. Slides were stained with aqueous solution of 1% Giemsa and coded for analysis. Abpout 100-200 well spread metaphase cells were scored per test point. Based on the observations made, the test chemical did not induce chromosome aberrations in CHEL cell line at dose level of 2 mM. It however induced gene mutation in CHEL cells in the presence and absence of S9 metabolic activation system above 2 mM.
- 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
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- Data is from study report
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Principles of method if other than guideline:
- The purpose of this study was to assess toxic and genotoxic effects of the test chemical on Chinese Hamster Ovary (CHO) cells by using several different in vitro-based assays, including genotoxicity tests based on the OECD Guideline No. 476 “In Vitro Mammalian Cell Gene Mutation Test”.
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- Cells deficient in hypoxanthine-guanine phosphoribosyl transferase (HPRT) due to the mutation HPRT+/- to HPRT-/- are resistant to cytotoxic effects of 6-thioguanine (TG). HPRT proficient cells are sensitive to TG (which causes inhibition of cellular metabolism and halts further cell division since HPRT enzyme activity is important for DNA synthesis), so mutant cells can proliferate in the presence of TG, while normal cells, containing hypoxanthine-guanine phosphoribosyl transferase cannot.
This in vitro test is an assay for the detection of forward gene mutations at the in hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus on the X chromosomes of hypodiploid, modal No. 20, CHO cells. Gene and chromosome mutations are considered as an initial step in the carcinogenic process.
The hypodiploid CHO cells are exposed to the test item with and without exogenous metabolic activation. Following an expression time the descendants of the treated cell population are monitored for the loss of functional HPRT enzyme.
HPRT catalyses the transformation of the purine analogues 6-thioguanine (TG) rendering them cytotoxic to normal cells. Hence, cells with mutations in the HPRT gene cannot phosphoribosylate the analogue and survive treatment with TG.
Therefore, mutated cells are able to proliferate in the presence of TG whereas the non-mutated cells die. However, the mutant phenotype requires a certain period of time before it is completely expressed. The phenotypic expression is achieved by allowing exponential growth of the cells for 7 days. - Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Ham's F-12K (Kaighn's) Medium containing 2 mM L-Glutamine supplemented with 10% Fetal Bovine Serum and 1% Penicillin-Streptomycin (10,000 U/mL).
- Properly maintained: Yes
- Periodically checked for Mycoplasma contamination: Not applicable
- Periodically checked for karyotype stability: Not applicable - Additional strain / cell type characteristics:
- other: Hypodiploid, modal No. 20
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 liver microsomal fraction obtained from Arcolor 1254-induced male Sprague-Dawley rats
- Test concentrations with justification for top dose:
- 0, 0.5, 1.0, 2.5 or 5.0 mM
- Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Water
Justification for choice of solvent/ vehicle: Polyethylene glycol was easily dissolved in water - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- Remarks:
- N-ethyl-N-nitrosourea (ENU) was the positive control substance in the tests done without S9
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: In medium with pre-incubation
DURATION
- Pre-incubation: One week involving 3 days of incubation with Hypoxanthine-aminopterin-thymidine (HAT) in medium as a mutant cleansing stage, followed by overnight incubation with hypoxanthine-thymidine (HT) in medium prior to a 3-4 days incubation in regular cell medium. After seeding and prior to treatment, the mutant-free cells were incubated for an additional of 24 hours.
- Exposure duration: 3 hours
- Expression time: 7 days
- Selection time: 14 days
- Fixation time: 7 days (harvest of cells)
SELECTION AGENT (mutation assays): 6-thioguanine (TG)
SPINDLE INHIBITOR (cytogenetic assays): Not applicable
STAIN (for cytogenetic assays): Crystal violet
NUMBER OF REPLICATIONS: A minimum of 2 replicates per dose concentration including negative and positive control.
NUMBER OF CELLS EVALUATED: 5 x 10 E5 cells were plated 7 days after treatment and whatever cells left, after 14 days of incubation with the selection medium, were evaluated.
DETERMINATION OF CYTOTOXICITY
- Method: After being exposed to the test chemical for 3 hours, in the absence or presence of S9, cells were trypsinized and 0.5 x 10 E5 cells per well was seeded in duplicates from two parallel duplicate cultures into 6-well plates in fresh medium. The relative total growth and cytotoxicity was evaluated 24 and 48 hours after seeding.
OTHER EXAMINATIONS: Not applicable
- Determination of polyploidy:
- Determination of endoreplication:
- Other:
OTHER: - Rationale for test conditions:
- No data
- Evaluation criteria:
- The cell line was observed for gene mutations
- Statistics:
- No data
- Species / strain:
- Chinese hamster Ovary (CHO)
- 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:
- No data
- Remarks on result:
- other: No mutagenic potential
- Conclusions:
- The test chemical in the concentration of 0, 0.5, 1.0, 2.5 or 5.0 mM does not cause genetic mutation(s) when CHO cells are exposed to the test chemical in the presence or abscence of metabolic activation.
- Executive summary:
An in vitro mammalian cell gene mutation study was designed and conducted to determine the genotoxicity profile of the test chemical when administered to Chinese Hamster Ovary (CHO) cells.
In the genotoxicity test, the test chemical was administered to CHO cells for 3 hrs at the dose levels of 0.5, 1.0, 2.5 or 5.0 mM and in the absence or presence of exogenous metabolic activation. CHO cells representing the negative controls were exposed to the vehicle. Positive controls, such as N-ethyl-N-nitrosourea (ENU) experiments without metabolic activation and 7,12-dimethylbenz(a) anthracene in experiments with metabolic activation, were also included in each test.
The positive control ENU showed indication of gene mutations occurring while no other treatment gave rise to gene toxicity.
When the mutation frequency was determined, a frequency of 3.79 x 10-4was shown after a 3 hour exposure of ENU as the positive control and in the absence of S9 liver microsomal fraction. Since no tested concentration of Polyethylene glycol, in the absence or presence of S9 liver microsomal fraction, resulted in colonies, we conclude that Polyethylene glycol does not give rise to gene mutations when CHO cells are exposed in vitro to the test chemical at 0, 0.5, 1.0, 2.5 or 5.0 mM for 3 hrs.
Conclusion
Based on the results of the current study, it is concluded that the test chemical does not give rise to gene mutations when CHO cells are exposed to the test chemical in vitro at 0, 0.5, 1.0, 2.5 or 5.0 mM for 3 hrs, in the presence or abscence of metabolic activation.
Referenceopen allclose all
Dose |
No Activation
(Negative) |
No Activation
(Negative) |
10% HLI
(Negative) |
10% HLI
(Negative) |
10% RLI
(Negative) |
10% RLI
(Negative) |
||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Protocol | Preincubation | Preincubation | Preincubation | Preincubation | Preincubation | Preincubation | ||||||
ug/Plate | Mean | ± SEM | Mean | ± SEM | Mean | ± SEM | Mean | ± SEM | Mean | ± SEM | Mean | ± SEM |
0 |
120 | 11.5 | 115 | 5.8 | 105 | 9.3 | 105 | 3 | 154 | 3.4 | 107 | 4.7 |
100 |
127 | 9 | 153 | 4.3 | 150 | 13.3 | 135 | 3.5 | 128 | 1.5 | 128 | 17.1 |
333 |
128 | 6 | 155 | 12.1 | 146 | 12 | 109 | 3.6 | 138 | 6.4 | 136 | 14.4 |
1000 |
132 | 3.7 | 150 | 8.1 | 143 | 6 | 140 | 10.1 | 125 | 5.5 | 116 | 6.4 |
3333 |
132 | 7.7 | 141 | 7.4 | 137 | 17.7 | 137 | 3.3 | 129 | 10.2 | 120 | 7.5 |
10000 |
119 | 5.3 | 141 | 9.5 | 137 | 5.4 | 127 | 2.2 | 155 | 4.8 | 137 | 7.6 |
Positive Control | 422 | 16.8 | 330 | 26 | 735 | 17.1 | 1359 | 38.4 | 445 | 37.5 | 526 | 25.3 |
Dose |
No Activation
(Negative) |
No Activation
(Negative) |
10% HLI
(Negative) |
10% HLI
(Negative) |
10% RLI
(Negative) |
10% RLI
(Negative) |
||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Protocol | Preincubation | Preincubation | Preincubation | Preincubation | Preincubation | Preincubation | ||||||
ug/Plate | Mean | ± SEM | Mean | ± SEM | Mean | ± SEM | Mean | ± SEM | Mean | ± SEM | Mean | ± SEM |
0 |
33 | 3.9 | 30 | 1.2 | 11 | 3.5 | 9 | 1.5 | 12 | 0.3 | 14 | 3.5 |
100 |
32 | 3.7 | 48 | 4.7 | 11 | 2.1 | 18 | 1.8 | 15 | 2.8 | 17 | 1.2 |
333 |
29 | 2 | 48 | 4.7 | 8 | 0.6 | 15 | 0.7 | 14 | 1.7 | 11 | 2 |
1000 |
29 | 7.2 | 47 | 1.3 | 16 | 1.2 | 17 | 0.7 | 13 | 3.3 | 13 | 2.2 |
3333 |
33 | 2.7 | 41 | 7.5 | 12 | 3.5 | 17 | 2.6 | 11 | 3.4 | 13 | 2.6 |
10000 |
32 | 2.6 | 54 | 7.7 | 12 | 2.6 | 17 | 0.7 | 14 | 2.3 | 6 | 1.9 |
Positive Control | 452 | 44.6 | 292 | 10 | 379 | 23.6 | 447 | 18.5 | 157 | 13 | 188 | 17.4 |
Table: Mutagenic effects of the test chemical
Dose (mM) |
n |
Aberrations/100 cells |
Aberrant cells |
Relative MI (%) |
|
Chromatid |
Chromosome |
||||
0 |
200 |
2.0 |
0.5 |
2.5 |
100 |
2.0 |
150 |
3.3 |
2.0 |
4.7 |
92 |
3.0 |
250 |
5.5 |
4.5 |
8.0b |
83 |
4.5 |
150 |
6.7 |
2.7 |
8.0a |
84 |
6.0 |
150 |
9.3 |
4.2 |
11.3a |
76 |
8.0 |
150 |
13.3 |
4.7 |
13.3a |
67 |
DMBA |
100 |
19.0 |
6.5 |
19.0a |
42 |
DMSO |
200 |
3.0 |
1.0 |
4.0 |
102 |
a Statistically significant at P<0.001 (Fischer’s exact test)
b Statistically significant at P<0.01 (Fischer’s exact test)
Table 1A. Effects of the test chemical exposure on gene toxicity in CHO cells. After being exposed to the test chemical for 3 hrs, cells was washed with sterile PBS and then incubated for 7 days at 37°C, 5% CO2. After 7 days, cells were re-seeded in new 6-well plates in the absence or presence of 10mM TG as a selection agent and returned to the incubator for 14 days at 37°C, 5% CO2. On day 15, all 6-well plates were stained with crystal violet and the number of colonies were counted manually. The results are presented as the total number of colonies found in the number of independent wells analyzed (e.g. 0 colonies in 4 wells will give 0/4) (n = 2 samples from 2 independent cultures).
|
With S9 |
Without S9 |
||
|
with TG |
without TG |
with TG |
without TG |
Neg. control |
0/4 |
593/4 |
0/4 |
667/4 |
Pos. control |
0/4 |
688/4 |
26/4 |
557/4 |
0.5 mM |
0/4 |
543/4 |
0/4 |
580/4 |
1.0 mM |
0/4 |
605/4 |
0/4 |
549/4 |
2.5 mM |
0/4 |
696/4 |
0/4 |
600/4 |
5.0 mM |
0/4 |
570/4 |
0/4 |
506/4 |
Table 1B. Mutation frequency in CHO cells after 3 hrs of exposure to test chemical in the absence or presence of 4% S9 liver microsomal fraction. N/A, no colonies present in the samples selected with TG, i.e. no mutation frequency could be determined.
|
With S9 |
Without S9 |
Neg. control |
N/A |
N/A |
Pos. control |
N/A |
3.79x10-4 |
0.5 mM |
N/A |
N/A |
1.0 mM |
N/A |
N/A |
2.5 mM |
N/A |
N/A |
5.0 mM |
N/A |
N/A |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Data available for the test chemical was reviewed to determine its mutagenic nature. The studies are as mentioned below:
Ames test:
Gene mutation toxicity study was performed for the test chemical to evaluate its mutagenic nature. The study was performed as per the preincubation protocol using Salmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system at doses of 0, 100, 333, 1000, 3333 or 10000 µg/plate. Water was used at the vehicle. The plates were incubated for 48 hrs after 20 mins preincubation before the evaluation of the revertant colonies could be made. Concurrent solvent and negative control chemicals were also included in the study. The test chemical did notinduce mutation in theSalmonella typhimurium strain TA100, TA1535, TA1537, TA98 both in the presence and absence of S9 metabolic activation system and hence the chemical is not likely to classify as a gene mutant in vitro.
In vitro mammalian chromosome aberration study:
In vitro mammalian chromosome aberration study was performed to determine the mutagenic nature of the test chemical. The study was performed using CHEL cellsin the presence and absence of S9 metabolic activation system. The test chemical was dissolved in DMSO and used at dose level of 0, 2, 3, 4.5, 6 or 8 mM. Concurrent solvent, untreated and positive control chemicals were also included in the study. 80000 CHEL cells were seeded in 25 cm2 flasks 9-20 hrs before treatment under standard culture conditions. The test chemical treatment was performed continuously for 16 or 24 hrs until harvesting. During the last 2 hrs of culture, colcemid was added. Cells were then collected by trypsin/EDTA treatment. Treated with hypotonic solution of tri-sodium citrate and washed twice with fresh fixative, dropped onto glass slides and air dried. Slides were stained with aqueous solution of 1% Giemsa and coded for analysis. Abpout 100-200 well spread metaphase cells were scored per test point. Based on the observations made, the test chemical did not induce chromosome aberrations in CHEL cell line at dose level of 2 mM. It however induced gene mutation in CHEL cells in the presence and absence of S9 metabolic activation system above 2 mM.
In vitro mammalian cell gene mutation assay:
An in vitro mammalian cell gene mutation study was designed and conducted to determine the genotoxicity profile of the test chemical when administered to Chinese Hamster Ovary (CHO) cells. In the genotoxicity test, the test chemical was administered to CHO cells for 3 hrs at the dose levels of 0.5, 1.0, 2.5 or 5.0 mM and in the absence or presence of exogenous metabolic activation. CHO cells representing the negative controls were exposed to the vehicle. Positive controls, such asN-ethyl-N-nitrosourea (ENU) experiments without metabolic activation and 7,12-dimethylbenz(a) anthracene in experiments with metabolic activation, were also included in each test. The positive control ENU showed indication of gene mutations occurring while no other treatment gave rise to gene toxicity. When the mutation frequency was determined, a frequency of 3.79 x 10-4was shown after a 3 hour exposure of ENU as the positive control and in the absence of S9 liver microsomal fraction. Since no tested concentration of Polyethylene glycol, in the absence or presence of S9 liver microsomal fraction, resulted in colonies, we conclude that Polyethylene glycol does not give rise to gene mutations when CHO cells are exposed in vitro to the test chemical at 0, 0.5, 1.0, 2.5 or 5.0 mM for 3 hrs.Based on the results of the current study, it is concluded that the test chemical does not give rise to gene mutations when CHO cells are exposed to the test chemical in vitro at 0, 0.5, 1.0, 2.5 or 5.0 mM for 3 hrs, in the presence or abscence of metabolic activation.
Based on the data available for the target chemical, the test chemical does not exhibit gene mutation in vitro. Hence the test chemical is not likely to be genetoxic in vitro as per the criteria mentoned in CLP regulation.
Justification for classification or non-classification
Based on the data available for the target chemical, the test chemical does not exhibit gene mutation in vitro. Hence the test chemical is not likely to be genetoxic in vitro as per the criteria mentoned in CLP regulation.
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