Amines, C10-14-tert-alkyl

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Substance is not genotoxic in vitro

Link to relevant study records

Reference

Reference 1

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:

other: Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Target gene:

HGPRT

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Details on mammalian cell type (if applicable):

- Type and identity of media: The cells were routinely maintained in Ham's F-12 nutrient mix (GIBCO, Grand Island, New York) supplemented with 5% (v/v) heat-inactivated (56°C, 30 minutes), dialyzed fetal bovine serum (GIBCO), antibiotics and antimycotics (penicillin G, 100 units/ml; streptomycin sulfate, 0.1 mg/ml; fungizone, 25 µg/ml; GIBCO) and an additional 2 mM L-glutamine (GIBCO).
The selection medium used for the detection of HGPRT- mutants was Ham's F-12 nutrient mix without hypoxanthine, supplemented with 10 µM 6 thioguanine (GIBCO) and 5% serum and the above-mentioned antibiotics.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes

Metabolic activation:

with and without

Metabolic activation system:

S9 liver homogenates prepared from Aroclor 1254-induced male Sprague-Dawley rats

Test concentrations with justification for top dose:

Preliminary Toxicity Assay
0 (solvent control), 3.13, 6.25, 12.5, 18.75, 25, 37.5, 50, 75, and 100 µg/ml

Initial Mutagenicity Assay
100, 110, and 120 µg/ml

Confirmatory Mutagenicity Assay
50 to 110 µg/ml in the absence of S9 and 50 to 160 µg/ml in the presence of S9

Repeat Confirmatory Mutagenicity Assay in the Presence of S9
0 (solvent control), 25, 50, 75, 100, 125, 150, 175, 200, 225, and 250 µg/ml

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: appropriate to dissolve the test material

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Ethyl methanesulfonate was used as the positive control for the non-activation system (without S9 factor). The positive control for assays performed with S9 (activation system) was 20-methylcholanthrene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: Cells in logarithmic growth phase were trypsinized and placed in medium containing 5% serum at a standard density of 3.0 x 106 cells/T-75 flask approximately 24 hours prior to treatment.
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 4 hours

STAIN (for cytogenetic assays): methanol/crystal violet

NUMBER OF REPLICATIONS: duplicate

NUMBER OF CELLS EVALUATED: 1 x 10e6 surviving cells

DETERMINATION OF CYTOTOXICITY
- Method: other: ability of the treated cells to form colonies

Evaluation criteria:

For an assay to be acceptable, the mutant frequency in positive controls should have been significantly higher than the solvent controls. An additional criteria, was that the mutant frequency in the solvent controls should have been within reasonable limits of the laboratory historical control values and literature values. The test chemical was considered positive if it induced a statistically significant, dose related, reproducible increase in mutant frequency. The final interpretation of the data took into consideration such factors as the mutant frequency and cloning efficiencies in the solvent controls.

Statistics:

The frequency of mutants per 106 clonable cells was statistically evaluated using a weighted analysis of variance; weights were derived from the inverse of the mutation frequency variance. The actual plate counts are assumed to follow a Poisson distribution therefore the mean plate count was used as an estimate of variance (Kirkland, 1989).
If the analysis of variance was significant at alpha = 0.05, a Dunnett's t-test was conducted (Winer, 1971), comparing each treated group and the positive control to the solvent control (alpha = 0.05, one-sided). Linear dose-related trend tests were performed if any of the pairwise comparisons of test material with the solvent control yielded significant differences.

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none
- Effects of osmolality: none


RANGE-FINDING/SCREENING STUDIES: The treated cultures in the absence of S9 showed toxicity with relative cell survival (RCS) values ranging from 7.7 to 108.6%. In the presence of S9 activation, moderate to no toxicity was observed with RCS values ranging from 34.1 to 102.0%. Based upon the results of this assay, concentration levels of 0 (solvent control), 25, 50, 70, 80, 90, 100, 110, and 120 µg/ml were selected for the initial gene mutation assay in the absence of S9 and 0 (solvent control), 25, 50, 80, 100, 110, 120, 130, and 140 µg/ml in the presence of S9.

COMPARISON WITH HISTORICAL CONTROL DATA: were within the range of the laboratory historical background

ADDITIONAL INFORMATION ON CYTOTOXICITY: In order to achieve the guideline stated acceptable level of cytotoxicity (10-20% survival compared to the solvent control) in the presence of S9 a third gene mutation assay was performed. Concentration levels in this assay were 0 (solvent control), 25, 50, 75, 100, 125, 150, 175, 200, 225, and 250 µg/ml.

Remarks on result:

other: all strains/cell types tested

Conclusions:

Interpretation of results:
negative

The results of the in vitro Chinese hamster ovary cell/hypoxanthine-guanine-phosphoribosyl transferase (CHO/HGPRT) forward gene mutation assay with the test material indicate that under the conditions of this study, the test article was non-mutagenic when evaluated in the absence or presence of an externally supplied metabolic activation (S9) system.

Executive summary:

The test material was evaluated in the in vitro Chinese hamster ovary cell/hypoxanthine-guanine-phosphoribosyl transferase (CHO/HGPRT) forward gene mutation assay. The genotoxic potential of the test material was assessed in two independent assays in the absence of an externally supplied metabolic activation (S9) and three independent assays in the presence of S9. The concentrations ranged from 25 to 120 µg/ml in the absence of S9 and from 25 to 250 µg/ml in the presence of S9. The highest concentration was based on the cytotoxicity of the test material to the cells. The adequacy of the experimental conditions for detection of induced mutation was confirmed by employing positive control chemicals, ethyl methanesulfonate for assays in the absence of S9 and 20-methylcholanthrene for assays in the presence of S9. Solvent control cultures were treated with the solvent used to dissolve the test material (i.e.ethanol). The results of thisin vitro CHO/HGPRT forward gene mutation assay with the test material indicate that under the conditions of this study, the test article was non-mutagenic when evaluated in the absence or presence of S9.

Reference 2

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Target gene:

not reported

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

Metabolic activation:

with and without

Metabolic activation system:

S-9 from rats induced with Aroclor 1254

Test concentrations with justification for top dose:

20 - 2000 ug/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: standard solvent

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Remarks:

2-anthramine, 2-nitrofluorene, sodium azide, and 9-aminoacridine

Positive control substance:

no

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
- Preincubation period: 10-hr
- Exposure duration: 72 hours
- Expression time (cells in growth medium): 10e8 to 10e9 cells per mL
- Selection time (if incubation with a selection agent): 72 hours



SELECTION AGENT (mutation assays): not reported
SPINDLE INHIBITOR (cytogenetic assays): not reported
STAIN (for cytogenetic assays): not reported


NUMBER OF REPLICATIONS: 3


DETERMINATION OF CYTOTOXICITY
- Method: elimination of a uniform background lawn

Evaluation criteria:

A mutagenicity assay is considered valid if the following conditions are met. First, the spontaneous reversion rate, with and without metabolic activation, must be reasonably consistent with the expected range for the strain being used. Second, the positive control materials must elicit a positive response. And third, strains must maintain characteristics, i.e., nutritional requirements, crystal violet sensitivity and ampicillin resistance. A test article is considered positive (mutagenic) if it elicits in independent assays a number of revertants per plate at least 2 times that observed in the solvent control (background). A response that does not meet this criteria but elicits a potential biologically significant response (e.g., a dose related increase in revertants per plate over 3 concentrations) is considered as equivocal response and requires further evaluation. A test article is considered negative (non-mutagenic) if the criteria for a positive assay were not met and the test article was tested up to 2,000 mg/plate or the limit of solubility or toxicity, whichever was lower. Toxicity is defined as the elimination of a uniform background lawn.

Statistics:

Statistical methods beyond the calculation of the mean and standard deviation are not considered necessary for the onterpretation of this study.

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

2000 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: the test article did precipitate
- Other confounding effects: none


RANGE-FINDING/SCREENING STUDIES: In 1984 (see IUCLID section 7.6.1, record IUC4#1), the test material was reported to be mutagenic in strain 1535. However, in that study, the test agent did not induce a dose related increase in mutation frequency. Rather, the sporadic positive response observed was based upon a statistical increase in mutation frequency compared to the negative controls. In contrast, our current criteria for a mutagenic response is based upon a two-fold or greater increase in mutation frequency compared to the negative controls. Because the marginal response observed in the 1984 study may not be biologically meaningful, the assay was repeated according to our current test system.


COMPARISON WITH HISTORICAL CONTROL DATA: not reported


ADDITIONAL INFORMATION ON CYTOTOXICITY: none

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'. Remarks: Salmonella typhimurium tester strains TA98, TA100, TA1535, TA1537

FREQUENCY OF EFFECTS: The test article did not induce an increase in revertants when compared to solvent controls.

Conclusions:

Negative

Executive summary:

The test material was evaluated for mutagenic activity in the Salmonella typhimurium gene mutation assay (Ames test). Tester strains were TA98, TA100, TA1535 and TA1537 in the presence and absence of a metabolic activation system (S-9 liver fraction from Aroclor 1254 induced rats). Dimethyl sulfoxide (DMSO) was used as the solvent for the test article and as the solvent control. In the presence of metabolic activation, 2 -anthramine was used as the positive control for all strains. In the absence of metabolic activation, the positive controls used were: 2 -nitrofluorene (TA98), sodium azide (TA100 and TA1535), and 9 -aminoacridine (TA1537). The test article was evaluated at concentrations ranging from 20 to 2000 ug/plate (all concentrations were based in the compound as received) and the number of revertants was determined.

The test article did not induce an increase in revertants when compared to solvent controls. This was true for all tester strains both with and without metabolic activation. Toxicity was observed in all strains, both without and with metabolic activation at 2000 ug/plate.

Under the conditions of this study, the test material is not mutagenic in the Salmonella gene mutation assay.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

Substance is not genotoxic in vivo in a guideline compliant mouse micronucleus assay

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

GLP compliance:

yes

Type of assay:

micronucleus assay

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Raleigh North Carolina, USA
- Age at study initiation: 8 weeks old
- Weight at study initiation: 22-31 g
- Assigned to test groups randomly: yes, computer randomization according to sex and body weight
- Fasting period before study: approximately 3 hours
- Housing: not reported
- Diet (e.g. ad libitum): ad libitum, Purina certified rodent chow 5002C
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 degrees C
- Humidity (%): 46-51%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark


IN-LIFE DATES: From: November 17, 1999 To: December 9, 1999

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: corn oil
- Justification for choice of solvent/vehicle: standard vehicle
- Concentration of test material in vehicle: none
- Amount of vehicle (if gavage or dermal): 10 ml/kg
- Type and concentration of dispersant aid (if powder): not applicable
- Lot/batch no. (if required): 92706
- Purity: not reported

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: 10 mL/kg


DIET PREPARATION not applicable

Duration of treatment / exposure:

single dose

Frequency of treatment:

single oral dose

Post exposure period:

2 days

Remarks:

Doses / Concentrations:
30, 150, 300 mg/kg
Basis:
nominal conc.

No. of animals per sex per dose:

5; Two additional animals per time point were dosed to account for possibility of unexpected deaths.

Control animals:

yes, concurrent vehicle

Positive control(s):

mitomycin C disolved in distilled water
- Justification for choice of positive control(s): standard
- Route of administration: intraperitoneal injection
- Doses / concentrations: 2.0 mg/kg

Tissues and cell types examined:

Clinical observations: recorded on days 0 and 1 and 2 post-dosing.
- Organs examined at necropsy: : bone marrow cells from both femurs were centrifuged, spread on glass slides, air-dried, fixed in methanol, stained with Acridine Orange stain and read with an epifluorescene microscope.
For each animal a total of at least 2000 polychromatic erythrocytes was scored for the presence or absence of micronuclei.

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: doses were selected after evaluating the results of an acute oral toxicity study in male and femae mice


TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): - Sampling times and number of samples: 24 and 48 hours post dosing. All animals from test groups and vehicle control (5/sex). Animals from the positive control groups (5/sex) at 24 hours after dosing.


DETAILS OF SLIDE PREPARATION: bone marrow cells from both femurs were centrifuged, spread on glass slides, air-dried for at least one hour, fixed in methanol fro 15 minutes and allowed to dry, stained with Acridine Orange stain and read with an epifluorescene microscope.


METHOD OF ANALYSIS: Slides from at least five animals per sex/dose group were observed when possible. Three slides were prepared per animal. Slides were coded and read blind in order to avoid bias on the part of the scorer. Slides were scanned for regios of suitable technical quality, where the cells were well spread, undamaged and well stained. These regions were normally located in a zone close to the middle of the smear. For each animal, a total of at least 1000 erythrocytes (polychromatic, referred to as PCE 1 and normochromatic) were recorded to calculate the PCE/NCE ratio. For each animal, the remaining number of polychromatic erythrocytes were recorded to total at least 2000 (referred to as PCED 2) and were scored for the presence or absence of micronuclei. Teh frequency of micronucleated polychromatic erythrocytes (MN-PCE) and the PCE/NCE ratio were calculated on the basis of these data.


OTHER: Cell counting was accomplished using the Xybion Path /Tox computer software system, G Module (GICELL program version 4.21) which captures data from the cell counter keyboard and provides an audit trail for quality assurance. Cell count data was entered, accepted and stored.

Evaluation criteria:

the test article is considered positive in this assay if it elicits a dose-response or a statistically significant increase in the number of micronucleated cells over that of the concurrent vehicle control at one or more dose levels. In the event that the test article elicits a significant increase in the number of MN-PCE due to an unusually low number of MN-PCE in the concurrent vehicle control, the data from that dose may be compared to historical vehicle control data. The test article is considered negative in this assay if: no indication of a dose-response is observed and the treatment groups do not show a statistically significant increase in the number of MN-PCE when compared to the vehicle control. - Criteria for selection of M.T.D.: the doses were selected after evaluation of the results of an acute oral toxicity in male and female CD-1 mice. The LD10 was selected as the high dose for this study since it was expected to produce significant toxicity, but not excessive lethality.

Statistics:

Data were analyzed separately for male and female animals using Statistical Analysis System (SAS, version 6.09 enhanced. An arcsine square root transformation was applied to the percent of micronucleated PCE's; all subsequent analyses for this parameter were conducted on transformed data. nitially, a three way analysis of variance model was applied to the data to determine the significance of each main effect and all two-way and three-way interaction effects. If significant interaction effects were identified, then the data were analyzed separately for each sex and/or day. Three independent single degree of freedom contrasts of the group means were used to test for trends in the group means and included in the assessment of 1) an overall effect of the test material treatment relative to control and 2) a linear dose-response trend among the tets material and 3) a quadratic dose-response trend among the test material. Additonally, pairwise comparisons between each of the three test material groups and the control group were made using Dunnett's t-test.

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Other: no range-finding study was conducted. Dose levels were selcted after evaluation of an acute oral toxicity study in mice.


RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): the test article did not induce an increase in the frequency of micronucleatedpolychromatic erythrocytes in bone marrow cells of male or female mice when compared to vehicle control values.
- Ratio of PCE/NCE (for Micronucleus assay): there was no statistically significant change in the polychromatic/normochromatic ratio at either 24 or 48 hours.
- Appropriateness of dose levels and route: Chosen after evaluating the results of an acute oral toxicity study in male and female mice
- Statistical evaluation: there was no statistically significant change in the polychromatic/normochromatic ratio at either 24 or 48 hours, which is indicative of the absence of cytotoxicity. An increase in the frequency of micronucleated polychromatic erythrocytes was observed in the bone marrow cells of male and female mice treated with 2.0 mg/kg of the positive control, mitomycin-C. When compared to the vehicle controls, the increase was greater than two-fold, indicating that the assay was sufficiently sensitive to detect induced cytogenetic damage. Statistical methods included analysis of variance followed by Dunnett’s T-Test on Least Square Means.

MORTALITY: there were no deaths in this study.
CLINICAL SIGNS: both male and female mice treated with 300 mg/kg of test article exhibited clinical signs of systemic
toxicity which included tremors and hyperactivity within 4 hours after treatment. Hyperactivity was also observed in
mice treated with 150 mg/kg of test article on day 0, after treatment. By day 1, all males exhibiting signs, with the
exception of one, and all females recovered.

		Number of animals exhibiting signs
Dose Group		Day 0	Day 1	Day 2
0 mg.kg Corn Oil
Male	Normal	10/10	10/10	5/5
Female	Normal	9/10	10/10	5/5
	Hyperactive	1/10	0/10	0/5
30 mg.kg Primene 81-R
Male	Normal	10/10	10/10	5/5
Female	Normal	10/10	10/10	5/5
150 mg.kg Primene 81-R
Male	Normal	9/10	10/10	5/5
	Hyperactive	1/10	0/10	0/5
Female	Normal	7/10	10/10	5/5
	Hyperactive	3/10	0/10	0/5
300 mg.kg Primene 81-R
Male	Normal	8/14	13/14	7/7
	Tremors	4/14	0/14	0/7
	Hyperactive	3/14	1/14	0/7
Female	Normal	5/14	14/14	7/7
	Tremors	1/14	0/14	0/7
	Hyperactive	7/14	0/14	0/7
	Respiratory Noise	1/14	0/14	0/7
2.0mg.kg Mitomycin C
Male	Normal	5/5	5/5	-
Female	Normal	5/5	5/5	-

Day 0 = Treatment Day

- No day 2 animals

DOSING SOLUTION ANALYSES: Samples of the dosing solutions were submitted for independent chemical analysis of the test article concentration. The results indicate that the concentration of the Primene 81R amine dosing solutions ranged from 96.2 to 76.8% of target value (highest to lowest concentrations respectively), which is within an acceptable range of the expected target concentration.

MICRONUCLEUS EVALUATION: The test article did not induce an increase i nthe frequency of micronucleated polychromatic erythrocytes in bone marrow cells of male or female mice when compared to the vehicle control values. This was true for both the 24 and 48 hour timepoints. There was no statistically significant change in the PCE/NCE ratio at either 24 or 48 hours, which is indicative of the absence of cytotoxicity.

An increase in the frequency of micronucleated polychromatic erythrocytes was observed in the bone marrow cells of male and female mice treated with 2.0 mg/kg of the positive control, mitomycin-C. When compared to the vehicle controls, the increase was greater than two-fold, indicating that the assay was sufficiently sensitive to detect induced cytogenetic damage.
EFFECT ON MITOTIC INDEX OR PCE/NCE RATIO: The test material was not mutagenic.

Conclusions:

Interpretation of results: negative
Under the condidtions of the study, the test material was not mutagenic in the micronucleus assay in CD-1 mouse marrow cells.

Executive summary:

The test material was evaluated for its potential to induce chromosomal damage in vivo, as assessed by the micronucleus assay with mouse bone marrow cells. Adult CD-1 male and female mice (5 male and 5 female animals per group, except for the high dose group, which had 2 additional animals per time point) received a single oral dose of the test article at concnetrations of 300, 150 or 30 mg/kg. Control animals received a single oral dose of corn oil (vehicle control), or an intraperitoneal injection of 2.0 mg/kg mitomycin-C (positve control) (MMC). Animals from test article and vehicle control groups were euthanized at 24 or 48 hours after treatment. Animlas from the positive control group were euthanized at 24 hours after treatment. Bone marrow slides were prepared and the frequency of micronucleated polychromatic erythrocytes was measured as an indicator of cytogenetic damage. For each animal, a total of at least 2000 polychromatic erythrocytes were scored for the presence or absence of micronuclei. In addition, the polychromatic erythrocyte/normochromatic erythrocyte (PCE/NCE) ratio was measured to evaluate the cytotoxicity of the test agent.

The test article did not induce an increase in the frquency of micronucleated polychromatic erythrocytes in bone marrow cells of males or female mice when compared to the vehicle controls. An increase in the frequency of micronucleated polychromatic erythrocytes was observed in the bone marrow cells of male and female mice trated with 2.0 mg/kg of the positive control, MMC. When compared to the vehicle controls, the increase was greater than two-fold, indicating that the assay was sufficiently sensitive to detect induced cytogenetic damge.

Under the condidtions of the study, the test material was not mutagenic in the micronucleus assay in CD-1 mouse marrow cells.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Mode of Action Analysis / Human Relevance Framework

Not applicable

Additional information

Short description of key information:
Genetic toxicity:
In vitro: Ames assay, bacterial cell- negative in all strains with and without metabolic activation (OECD TG 471)
in vitro: Gene mutation, mammalian cell- negative in CHO cells with and without metabolic activation (OECD TG 476)
In vivo: Mouse micronucleus assay- negative (OECD TG 474)

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

The test material was negative in both in vitro and in vivo genetic toxicity testing. There is no need for classification.