Registration Dossier
Registration Dossier
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EC number: 700-111-0 | CAS number: 148528-05-6
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Link to relevant study records
- Endpoint:
- genetic toxicity in vitro, other
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- other: The predicted compound is outside the Applicability Domain of the model
- Justification for type of information:
- QSAR prediction
- Principles of method if other than guideline:
- Mutagenicity (Ames test) model (CAESAR) 2.1.13
Mutagenicity (Ames test) model (SarPy/IRFMN) 1.0.7
Mutagenicity (Ames test) model (ISS) 1.0.2
Mutagenicity (Ames test) model (KNN/Read-Across) 1.0.0 - GLP compliance:
- no
- Remarks:
- not applicable
- Type of assay:
- other: QSAR
- Species / strain:
- other: not applicable
- Metabolic activation:
- not applicable
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: not applicable
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- not applicable
- Remarks on result:
- no mutagenic potential (based on QSAR/QSPR prediction)
- Conclusions:
- Interpretation of results: negative
With the models (CAESAR) 2.1.13, (ISS) 1.0.2 and (KNN/Read-Across) 1.0.0 the mutagen activity of the substance was predicted as non-mutagen. With the model (SarPy/IRFMN) 1.0.7 the mutagen activity of the substance was predicted as possible non-mutagen. - Endpoint:
- genetic toxicity in vitro, other
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Model considered reliable by OECD
- Justification for type of information:
- QSAR prediction
- Qualifier:
- according to guideline
- Guideline:
- other: Guidance Document on the Validation of (Quantitative) Structure-Activity relationship models (OECD, 2007) and Guidance on Information Requirements and Chemical Safety Assessment/Chapter R.6: QSARS and Grouping Chemicals (ECHA, 2008)
- Principles of method if other than guideline:
- Read-across from category members
- GLP compliance:
- no
- Remarks:
- not applicable
- Type of assay:
- other: QSAR
- Species / strain:
- other: not applicable
- Metabolic activation:
- not applicable
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: not applicable
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- not applicable
- Remarks on result:
- no mutagenic potential (based on QSAR/QSPR prediction)
- Conclusions:
- Interpretation of results: negative
- Executive summary:
The read-across models predict this material is not mutagenic in salmonella typhimurium reverse
mutagenic (Ames) testing.
- Endpoint:
- genetic toxicity in vitro, other
- Type of information:
- (Q)SAR
- Adequacy of study:
- weight of evidence
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- other: The official QMRF and QPRF formats were not met
- Justification for type of information:
- QSAR prediction
- Principles of method if other than guideline:
- Consensus method
- GLP compliance:
- no
- Remarks:
- not applicable
- Type of assay:
- other: QSAR
- Species / strain:
- other: not applicable
- Metabolic activation:
- not applicable
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- other: not applicable
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- not applicable
- Remarks on result:
- no mutagenic potential (based on QSAR/QSPR prediction)
- Conclusions:
- Interpretation of results: negative
Mutagenicity 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:
- weight of evidence
- Study period:
- 28 April 1997 to 28 July 1997
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: The study was conducted according to GLP. Cytotoxicity was only evaluated during a range finding study.
- Qualifier:
- according to guideline
- Guideline:
- other: (Kanpogyo Notification No. 39, Yakuhatsu Notification No. 229, jointly issued with 59 Kikyoku Notification No. 85
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: About Standard for Investigation by Chromosome Aberration Test using Cultured Mammalian Cells”(Kihatsu No. 143 issued by the director general of Ministry of Labor’s Labor Standards Bureau on March 18, 1987
- Deviations:
- not specified
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- mammalian cell line, other: The fibroblast cell line, CHL/IU, derived from the lungs of newborn female Chinese hamsters.
- Details on mammalian cell type (if applicable):
- - Type and identity of media:
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: Not reported
- Periodically checked for karyotype stability: Not reported
- Periodically "cleansed" against high spontaneous background: yes - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9
- Test concentrations with justification for top dose:
- The presence or absence of structural and numerical aberrations was determined at 12.5, 25, 50, and 100 µg/ml in the absence of metabolic
activation with the treatment periods of 24 and 48 hrs, at 44.4, 66.7, 100, and 150 µg/ml in the presence of metabolic activation without S9 Mix, and
at 444, 667, 1,000, and 1,500 µg/ml in the presence of metabolic activation with S9 Mix. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: physiological saline and DMSO
- Justification for choice of vehicle: solubility - Untreated negative controls:
- yes
- Remarks:
- Physiological saline
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- Migrated to IUCLID6: MMC; Lot Number ESH5706; biochemical grade
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- Migrated to IUCLID6: Lot No. ECF6942; purity, 98%; analytical grade
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: agar plate (plating method)
DURATION
- Preincubation period: not applicable
- Exposure duration:
Dose-ranging: 24 and 48 hours (direct method), 24 hours (metabolic activation method)
Main study: 24 and 48 hours (direct method), 6 hours (metabolic activation method)
- Expression time (cells in growth medium): 72 hours
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells):
Dose-ranging: 24 and 48 hours (direct method), 24 hours (metabolic activation method)
Main study: 24 and 48 hours (direct method), 24 hours (metabolic activation method)
SELECTION AGENT (mutation assays): not used
SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): 3.0% Giemsa stain
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 100 per replicate
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: growth inhibition
OTHER EXAMINATIONS:
- Determination of polyploidy: Structural aberrations were classified into chromatid and chromosome gaps (gaps), chromatid break (ctb),
chromatid exchange (cte), chromosome break (csb), chromosome exchange (cse), and others (o) according to the Chromosomal Aberration Atlas6).
- Determination of endoreplication:
- Other:
OTHER: - Statistics:
- The incidence of cells with chromosomal aberrations at each dose was analyzed and compared between the negative control group and each test
substance group by the ¿2-test. If significant differences were found, the binominal test was conducted between the background data from the
negative control group and data from each test substance group. If significant differences were observed, Cochran-Armitage’s trend test was
conducted to determine the presence or absence of dose-dependency. The test result was judged to be positive if all these tests showed significant
differences. Otherwise, the test result was regarded as negative. The D20 value was calculated if the test result was positive. Data obtained from
the positive control group were subjected to the ¿2-test and binominal test, and the test result was regarded as positive if both tests showed
significant differences. - Species / strain:
- other: The fibroblast cell line, CHL/IU, derived from the lungs of newborn female Chinese hamsters.
- Metabolic activation:
- without
- Genotoxicity:
- ambiguous
- Cytotoxicity / choice of top concentrations:
- not determined
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- other: The fibroblast cell line, CHL/IU, derived from the lungs of newborn female Chinese hamsters.
- Metabolic activation:
- with
- Genotoxicity:
- ambiguous
- Cytotoxicity / choice of top concentrations:
- not determined
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: Not reported
- Effects of osmolality: Not reported
- Evaporation from medium: Not reported
- Water solubility:
- Precipitation:
- Other confounding effects:
RANGE-FINDING/SCREENING STUDIES:
A dose-ranging study was undertaken according to the testing guidelines in the presence of metabolic activation (using S9 Mix with a treatment
period of 6 hrs and a recovery period of 18 hrs and without using S9 Mix) and the absence of metabolic activation (with a treatment period of 24 or 48hrs) in order to select doses of the test substance for the main study by determining whether or not cell growth was inhibited2)3). The highest dose of the test substance was set at 5,000 µg/mL, followed by 1,500, 500, 150, 50, 15, and 5 µg/mL with a common ratio of about 3. The test
substance and negative control (physiological saline (JP) used as solvent) were used.
The results of the range finding study are tabulated in the "Any other information on results incl. tables" section below.
- Conclusions:
- Interpretation of results:
positive with metabolic activation
positive without metabolic activation
It was concluded that MODA inhibited cell division regardless of the presence or absence of S9 Mix and induced structural chromosomal aberrations in the absence of S9 Mix under the conditions of the present study. The responses were generated at highly cytotoxic doses and should be interpreted with caution. Therefore a new in vitro micronucleus study (OECD 487, 2015) was conducted with the aim to evaluate the genotoxicity in an appropriate concentration range and with the determination of the cytotoxicity simultaneously during the main study. - Executive summary:
- A study was conducted at Shin Nippon Biomedical Laboratories, Ltd, Japan, in order to evaluate whether or not the substance 2-methyl-1,8-octanediamine (MODA) induces chromosomal aberrations for the Chinese hamster's lung-derived fibroblast cell line CHL/IU. The study was conducted to GLP and according to the Japenese Guidelines 'Kanpogyo Notification No. 39, Yakuhatsu Notification No. 229, jointly issued with 59 Kikyoku Notification No. 85' and to 'Kihatsu No. 143'. The presence or absence of structural and numerical aberrations was determined at 12.5, 25, 50, and 100 µg/mL in the absence of metabolic activation with the treatment periods of 24 and 48 hrs, at 44.4, 66.7, 100, and 150 µg/mL in the presence of metabolic activation without S9 Mix, and at 444, 667, 1,000, and 1,500 µg/mL in the presence of metabolic activation with S9 Mix. The incidence of cells with structural aberrations significantly increased compared with the negative control in the absence of metabolic activation with the treatment periods of 24 and 48 hrs, as well as in the presence of metabolic activation without S9 Mix. The incidence of cells with numerical aberrations significantly increased compared to the negative control at 44.4-100 µg/mL in the presence of metabolic activation without S9 Mix and at 1,000 µg/mL in the presence of metabolic activation with S9 Mix. In addition, C-mitoses, which are not considered as aberrations but are thought to be due to the inhibition of spindle formation, were observed in high dose groups in the absence of metabolic activation and mid to high dose groups in the presence of metabolic activation without S9 Mix. The incidence of cells with structural and numerical aberrations in the negative and positive control groups was within the standard ranges of this laboratory. From these findings, it was concluded that MODA inhibited cell division regardless of the presence or absence of S9 Mix and induced structural chromosomal aberrations in the absence of S9 Mix under the conditions of the present study.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 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)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- Thymidine Kinase (TK)
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: complete growth medium F10P [Dulbecco’s modified Eagle’s Medium (DMEM) with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplied with 0.1 % Pluronic, 90 % and Fetal bovine serum, 1 0%]. Treatment medium F5P [Dulbecco’s modified Eagle’s Medium (DMEM) with 4 mM Lglutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplied with 0.1 % Pluronic, 95 % and Fetal bovine serum, 5 %].
- Metabolic activation:
- with and without
- Metabolic activation system:
- A co-factor supplemented postmitochondrial fraction (S9) from the liver of Aroclor 1254 induced Sprague- Dawley rats.
- Test concentrations with justification for top dose:
- 2.5 µg/ml, 1.25 µg/ml, 0.625 µg/ml and 0.3125 µg/ml
- Vehicle / solvent:
- none
- Untreated negative controls:
- yes
- Remarks:
- sterile distilled water
- Negative solvent / vehicle controls:
- no
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- methylmethanesulfonate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): 13 days
SELECTION AGENT (mutation assays): Trifluorothymidine (TFT)
NUMBER OF REPLICATIONS: 3
NUMBER OF CELLS EVALUATED: 3 x 10e6
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth - Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Remarks:
- Cytotoxic effect of cell growth inhibition was observed at 2.5 µg/ml and above.
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: high pH values (pH 11.31 (0.6 mg/mL-water); pH 11.72 (3 mg/mL-water); pH 12.10 (15 mg/mL-water)) result in cytotoxicity
- Precipitation: none
RANGE-FINDING/SCREENING STUDIES: The testing concentrations of the test substances for the study were selected based on the results of preliminary range-finding test using L5178Y/TK+/- mouse lymphoma cells. In the range-finding test, cytotoxic effect of cell growth inhibition was observed at 2.5 µg/ml and above. - Conclusions:
- Interpretation of results: negative
From the above study, the test item- methyl-1,8-octanediamine (MODA) is non-mutagenic to the L5178Y TK+/- clone, both in the absence and presence of S9 metabolic activation system. - Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 28 April 1997 to 14 July 1997
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study conducted according to the principles of GLP.
- Qualifier:
- according to guideline
- Guideline:
- other: the Chemical Substances Control Law Test Guidelines (“Amendments of the ‘Test Methods for New Chemical Substances’”) (Kanpogyo Notification No. 700, Yakuhatsu Notification No. 1039; issued jointly with 61 Kikyoku Notification No. 1014
- Deviations:
- not specified
- Qualifier:
- according to guideline
- Guideline:
- other: Industrial Safety and Health Act Test Guidelines (Criteria Determined by the Ministry of Labor in Accordance with Paragraph 1, Article 57-2 of the Industrial Safety and Health Law (Act No. 57 of 1972) (Ministry of Labor Notice No. 77 of September 1, 1988)
- Deviations:
- not specified
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- GLP compliance:
- yes
- Type of assay:
- bacterial gene mutation assay
- Target gene:
- Not reported
- 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:
- S9
- Test concentrations with justification for top dose:
- Based on the findings of a dose ranging study, the test substance concentrations were set at 156, 313, 625, 1250, 2500 and 5000 µg/plate both in
the presence and absence of metabolic activation. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water for injection (batch No. 6C73N; supplied by Otsuka Pharmaceutical Factory Inc.)
- Justification for choice of solvent/vehicle: solubility - Untreated negative controls:
- yes
- Remarks:
- water for injection
- Positive controls:
- yes
- Positive control substance:
- furylfuramide
- Remarks:
- (Lot No.: SAJ0748; purity: 99.4%; analytical grade; Wako Pure Chemical Industries, Ltd.)
- Positive controls:
- yes
- Positive control substance:
- N-ethyl-N-nitro-N-nitrosoguanidine
- Remarks:
- (Lot No.: V1A8648; purity: 97%; analytical grade; Nacalai Tesque, Inc.)
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- (Lot No.: M0T3555; purity: 95%; analytical grade; Nacalai Tesque, Inc.)
- Positive controls:
- yes
- Positive control substance:
- other: 2-Aminoanthracene
- Remarks:
- (Lot No.: DSR3205; purity: 92.6%; analytical grade; Wako Pure Chemical Industries, Ltd.)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Preincubation period: 12 hours (pre-culture)
- Exposure duration: 48 hours
- Expression time (cells in growth medium): not applicable
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): not applicable
SELECTION AGENT (mutation assays): not applicable
SPINDLE INHIBITOR (cytogenetic assays): not applicable
STAIN (for cytogenetic assays): not applicable
NUMBER OF REPLICATIONS: not applicable
NUMBER OF CELLS EVALUATED: not applicable
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: growth inhibition
OTHER EXAMINATIONS:
- Determination of polyploidy: not applicable
- Determination of endoreplication: not applicable
- Other: not applicable
- Evaluation criteria:
- The test substance was regarded as mutagenic if the (mean) revertant colony count per plate showed 2-folds or greater increases compared to the
negative control, if these increases were dose-dependent, and if reproducibility is noted in the dose-ranging study and the main study. Otherwise,
the test substance was considered to be non-mutagenic. - Statistics:
- No statistical analysis was conducted in the present study.
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 2,500 - 5,000 µg/plate
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 2,500 - 5,000 µg/plate
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 2,500 - 5,000 µg/plate
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 2,500 - 5,000 µg/plate
- Untreated negative controls validity:
- valid
- 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
- Remarks:
- at 5000 µg/plate
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: Not reported
- Effects of osmolality: Not reported
- Evaporation from medium: Not reported
- Water solubility: Not reported
- Precipitation: The test substance did not precipitate up to 5000 µg/plate regardless of the presence or absence of metabolic activation.
- Other confounding effects: None
RANGE-FINDING/SCREENING STUDIES:
Negative with and without metabolic activation
COMPARISON WITH HISTORICAL CONTROL DATA:
In both the dose-ranging study and the main study, revertant colony counts noted in the negative and positive control groups were within the normalrange
ADDITIONAL INFORMATION ON CYTOTOXICITY: None - Conclusions:
- Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation
It was concluded that MODA does not show mutagenic potential under the conditions of the present study regardless of the presence or absence of metabolic activation. - Executive summary:
A reverse mutation study of 2-methyl-1,8-octanediamine (MODA) was conducted at Shin Nippon Biomedical Laboratories, Ltd., Japan, by the plate method using the following five strains in order to evaluate the mutagenic potential of MODA: Salmonella typhimurium TA98, TA100, TA1535 and TA1537 and Escherichia coli WP2urvA. The study was conducted according to the Japanese Guidelines:
i. Kanpogyo Notification No. 700, Yakuhatsu Notification No. 1039, (issued jointly with 61 Kikyoku Notification No. 1014) and
ii. Article 57-2 of the Industrial Safety and Health Law (Act No. 57 of 1972) (Ministry of Labor Notice No. 77 of September 1, 1988).
The study was also conducted according to the principles of GLP. Seven different doses of the test substance, ranging between 5 and 5,000 µg/plate with a common ratio of about 3, were used in the dose-ranging study regardless of whether metabolic activation was conducted. Six different doses of the test substance, ranging between 156 and 5,000 µg/plate with a common ratio of about 2, were used in the main study regardless of whether metabolic activation was conducted. MODA did not cause 2-folds or greater increases in the revertant colony count for any of the five strains compared to the negative control regardless of the presence or absence of metabolic activation in either the dose-ranging study or the main study. In the absence of metabolic activation, the growth of TA100, TA1535 and TA1537 was inhibited at 2,500 µg/plate or higher concentrations, while that of WP2urvA and TA98 was inhibited at 5,000 µg/plate.
In the presence of metabolic activation, the growth of TA100, TA1535, TA98, and TA1537 was inhibited at 5,000 µg/plate. The test substance did not precipitate up to 5,000 µg/plate regardless of the presence or absence of metabolic activation.
The number of revertant colonies was within the reference range of this laboratory in both the negative control group and positive control groups, indicating that this study was conducted under appropriate conditions. Based on these findings, it was concluded that MODA does not show mutagenic potential under the conditions of the present study regardless of the presence or absence of metabolic activation.
- Endpoint:
- in vitro cytogenicity / micronucleus study
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 2015-01-19 to 2015-03-24
- Reliability:
- 1 (reliable without restriction)
- Qualifier:
- according to guideline
- Guideline:
- other: OECD 487 in vitro mammalian cell micronucleus test (26 September 2014)
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- in vitro mammalian cell micronucleus test
- Species / strain / cell type:
- lymphocytes: human peripheral blood lymphocytes
- Details on mammalian cell type (if applicable):
- - Blood samples were obtained by venapuncture from young healthy, non-smoking individuals (24 and 34 years old) with no known recent exposures to genotoxic chemicals or radiation
- Type and identity of media: The medium for culturing the human peripheral blood lymphocytes consisted of RPMI 1640 medium (with HEPES and Glutamax), supplemented with heatinactivated (30 min, 56ºC) foetal calf serum (20%), penicillin (100 U/ml medium), streptomycin (100 µg/ml medium) and phytohaemagglutinin (2.4 µg/ml) - Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Experiment 1: 1266, 633, 317, 158, 79, 40, 20, 10, 5, 2.5 µg/mL (pulse treatment method with and without metabolic activation)
Experiment 2: 1266, 886, 620, 434, 303, 213 µg/mL (pulse treatment method without metabolic activation)
Experiment 2: 1266, 886, 620, 434, 303, 213, 149, 104, 72.8, 51.1, 35.8, 25.0, 17.5, 12.3 µg/mL (continuous treatment method without metabolic activation) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: RPMI 1640 medium
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- culture medium
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- other: vinblastine sulphate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: experiment 1 and 2: 48 hours
- Exposure duration and Expression time: experiment 1: 4 hours treatment time and 20 hours recovery time; experiment 2: 24 hours treatment time
- Selection time (if incubation with a selection agent): experiment 1 and experiment 2 (pulse treatment method): 20 hours (during recovery time); experiment 2 (continuous treatment method): 24 hours (during treatment time)
- Fixation time (start of exposure up to fixation or harvest of cells): 72 hours (at the end of the total incubation period)
SELECTION AGENT (mutation assays): cytochalasin B
STAIN (for cytogenetic assays): acridin-orange
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 500 cells per slide (in total 1000 cells per dose level)
DETERMINATION OF CYTOTOXICITY
- Method: Cytokinesis-Block Proliferation Index (CBPI) - Evaluation criteria:
- The frequencies of binucleated cells with micronuclei were used for the evaluation of micronuclei induction. The CBPI was calculated for treated (selected doses) and control cultures as a measure of cell cycle delay. If observed, the concurrent measures of cytotoxicity (cell density on the slides, signs of apoptosis or necrosis) were recorded for all treated and negative control cultures.
The study was considered valid if the clastogenic and aneugenic positive controls gave a statistically significant increase in the number of binucleated cells containing micronuclei and if the negative controls (culture medium) were within the historical data performed at the test facility.
A response was considered positive if a statistically significant concentration-related or a reproducible statistically significant increase in the number of binucleated cells containing micronuclei was induced, at any of the test points. The biological relevance will be considered first. During evaluation, the observed values will be compared with the historical control range. Statistical methods will be used as an aid in evaluating the test results but will not be the only determining factor for a positive response. However, the results of statistical testing will be assessed with respect to dose-response relationship.
A response was considered equivocal if the percentage of binucleated cells containing micronuclei was marginally statistically significant higher than that of the negative control (0.05 - Statistics:
- Chi-square test (one-sided):
The frequencies of micronuclei found in the cultures treated with the test substance and positive control cultures were compared with those of the concurrent negative control using the Chi-square test (one-sided). The results were considered statistically significant when the p-value of the Chi-square test was less than 0.05. - Species / strain:
- lymphocytes: human peripheral blood lymphocytes
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: up to the testet concentration of 1266 µg/mL the pH of the cultures was not changed by more than 1 pH unit when compared to the solvent control.
- Cultures were slightly haemolytic at the concentration of 1266 µg/mL in experiment 1 without metabolic actiovation and experiment 2 continuous treatment without metabolic activation
COMPARISON WITH HISTORICAL CONTROL DATA: yes
ADDITIONAL INFORMATION ON CYTOTOXICITY:
- Due to a steep concentration response curve for the cytotoxicity of the test substance, concentrations achieving the moderate toxicity level were not obtained and therefore this group was repeated in the second experiment simultaneously with the continuous treatment group.
- In the pulse treatment groups both with and without metabolic activation (S9-mix), at the highest concentration (1266 µg/ml) a low cell density on the slides was observed indicating cell death in the cultures. As a result, the calculated cytotoxicity from the CBPI values may be an underestimation of the actual cytotoxicity
- In the second experiment, in the pulse treatment group without metabolic activation, at the highest concentration of 1266 µg/ml, the cell density on the slides was very low. The calculated cytotoxicity of 1% was therefore considered an underestimation of the actual cytotoxicity - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative
From the results obtained in the in vitro micronucleus test it is concluded that, under the conditions used in this study, the test substance, MODA, was not clastogenic and/or aneugenic to cultured human lymphocytes.
Referenceopen allclose all
Direct method |
24-hr treatment |
Dose (µg/mL) |
0 |
5 |
15 |
50 |
150 |
500 |
1,500 |
5,000 |
Cell growth (%)* |
100 |
102.5 |
84 |
40 |
12 |
0 |
0 |
0 |
||
48-hr treatment |
Dose (µg/mL) |
0 |
5 |
15 |
50 |
150 |
500 |
1,500 |
5,000 |
|
Cell growth (%)* |
100 |
82 |
76 |
35 |
7 |
0 |
0 |
0 |
||
Metabolic activation method |
Without S9 Mix |
Dose (µg/mL) |
0 |
5 |
15 |
50 |
150 |
500 |
1,500 |
5,000 |
Cell growth (%)* |
100 |
118 |
112 |
74 |
21 |
0 |
0 |
0 |
||
With S9Mix |
Dose (µg/mL) |
0 |
5 |
15 |
50 |
150 |
500 |
1,500 |
5,000 |
|
Cell growth (%)* |
100 |
99 |
96.5 |
94 |
91 |
64 |
17 |
0 |
MODA did not cause 2-folds or greater increases in the revertant colony count for any of the five strains compared to the negative control regardless of the presence or absence of metabolic activation in either the dose-ranging study or the main study. The number of revertant colonies was within the reference range of this laboratory in both the negative control group and positive control groups, indicating that this study was conducted under appropriate conditions.
Table 1. Results of the Main Study
Metabolic activation |
Concentration of test substance (µg/plate) |
Revertant colony counts (per plate) |
|||||
Base-pair substitution |
Frame shift |
||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
|||
S9 Mix (-) |
Negative control (water for injection) |
134, 136 (135) |
10, 10 (10) |
23, 17 (20) |
17, 16 (17) |
7, 10 (9) |
|
156 |
142, 147 (145) |
11, 7 (9) |
22, 23 (23) |
23, 21 (22) |
6, 13 (10) |
||
313 |
135, 125 (130) |
16, 15 (16) |
24, 27 (26) |
28, 24 (26) |
9, 12 (11) |
||
625 |
134, 123 (129) |
16, 13 (15) |
22, 17 (20) |
23, 17 (20) |
4, 8 (6) |
||
1250 |
138, 123 (131) |
15, 13 (14) |
26, 20 (23) |
19, 28 (24) |
10, 12 (11) |
||
2500 |
83*, 97* (90) |
3*, 6* (5) |
23, 23 (23) |
18, 16 (17) |
5*, 4* (5) |
||
5000 |
0***, 0** (0) |
0**, 0** (0) |
13*, 15* (14) |
0**, 0** (0) |
0**, 0** (0) |
||
S9 Mix (+) |
Negative control (water for injection |
141, 150 (146) |
11, 13 (12) |
20, 20 (20) |
38, 33 (36) |
14, 10 (12) |
|
156 |
127, 125 (126) |
8, 10 (9) |
18, 20 (19) |
23, 34 (29) |
9, 11 (10) |
||
313 |
127, 124 (126) |
7, 9 (8) |
21, 18 (20) |
34, 32 (33) |
5, 7 (6) |
||
625 |
138, 127 (133) |
10, 13 (12) |
20, 22 (21) |
35, 31 (33) |
7, 14 (11) |
||
1250 |
128, 116 (122) |
13, 14 (14) |
31, 18 (25) |
36, 30 (33) |
12, 9 (11) |
||
2500 |
96, 113 (105) |
18, 11 (15) |
26, 25 (26) |
22, 33 (28) |
13, 10 (12) |
||
5000 |
57*, 64* (61) |
0**, 0** (0) |
15, 30 (23) |
1*, 0** (1) |
4*, 5* (5) |
||
Positive control |
Without S9 Mix |
Name |
AF-2 |
ENNG |
ENNG |
AF-2 |
9AA |
Concentration (µg/plate) |
0.01 |
5 |
2 |
0.1 |
80 |
||
Colony counts/plate |
477, 432 (455) |
187, 191 (189) |
568, 537 (553) |
685, 657 (671) |
364, 458 (411) |
||
With S9 Mix |
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
|
Concentration (µg/plate) |
1 |
2 |
10 |
0.5 |
2 |
||
Colony counts/plate |
1886, 1861 (1874) |
445, 450 (448) |
559, 514 (537) |
934, 782 (858) |
616, 550 (583) |
||
(Remarks)
1. Figures in parentheses: means of two plates
2. Names of positive controls
AF-2: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide
ENNG: N-ethyl-N'-nitro-N-nitrosoguanidine
9AA: 9-aminoacridine
2AA: 2-aminoanthracene
3.* : The growth of background bacteria on the plate was apparently inhibited compared to the negative control.
**: The growth of background bacteria on the plate was inhibited by the test substance, resulting in their death with
small colonies as observed with the naked eye.
***: The growth of background bacteria on the plate was strongly inhibited by the test substance, resulting in their death and the absence of small colonies.
- Micronuclei induction as a result of treatment with the negative control and the positive controls:
In both the first and second experiment, the negative control (culture medium) was within the range of historical data of the test facility, except for the continuous treatment group, were the number of micronuclei found in the binucleated cells was slightly below the lower end of the historical range. As the observed low frequency of the micronuclei in this group may be the result of inter individual variation in the blood donors and the result is only marginally below the range, the experiment is still considered valid.
- Micronuclei induction as a result of treatment with the test substance: In the first experiment, in the pulse treatment groups both with and without
metabolic activation, three dose levels of the test substance (1266, 633 and 317 µg/mL), together with the solvent control (culture medium) and
positive control, were analyzed for micronucleus induction in binucleated lymphocytes.
In the pulse treatment group with metabolic activation, the highest concentration of the test substance resulted in a statistically marginally higher (p = 0.0841) increase in the number of binucleated cells containing micronuclei, when compared to the numbers found in the concurrent control cultures (Table 1). However, the number of binucleated cells containing micronuclei for this concentration was equal to the upper level of the historical control range and considering the fact that this concentration induced an increase in pH of (almost) one unit, this result is considered to be a matter of chance and not biologically relevant.
In the pulse treatment group without metabolic activation, the test substance did not show a statistically significant increase in the number of binucleated cells containing micronuclei, at any of the concentrations analysed when compared to the numbers found in the concurrent control cultures (Table 2).
In the second experiment, in the pulse treatment group without metabolic activation, the highest concentration (1266 µg/ml) was not selected for analysis of micronuclei induction in binucleated cells due to the low cell density on the slides (indicating severe cytotoxicity). Three dose levels of the test substance (886, 434 and 213 µg/mL), together with the solvent control (culture medium), were analyzed for micronucleus induction in binucleated lymphocytes.
In the continuous treatment group without metabolic activation, at the concentration of 886 µg/mL, the observed cytotoxicity (65%) was slightly higher than the cytotoxicity (55 ± 5%) as defined in the study plan. Nevertheless, the observed cytotoxicity was close enough to the defined range to be considered acceptable. Therefore, four dose levels of the test substance (886, 620, 303 and 104 µg/mL), together with the solvent control (culture medium) and positive control were analysed for micronucleus induction in binucleated lymphocytes.
In the second experiment, in the pulse treatment group without metabolic activation, the highest concentration (886 µg/mL) of the test substance resulted in a statistically marginally higher (p = 0.0577) increase in the number of binucleated cells containing micronuclei, when compared to the numbers found in the concurrent control cultures (Table 3). However, the number of binucleated cells containing micronuclei for this concentration was well within the historical control range. In addition, in the first experiment in the pulse treatment group without metabolic activation, no increase was observed when tested up to the highest feasible concentration of 1266 µg/mL. Hence, this result is not reproducible and thus considered to be a matter of chance and not biologically relevant.
In the continuous treatment group without metabolic activation, at the lowest concentration analysed (104 µg/mL), the test substance showed a statistically marginally higher (p = 0.0537) increase in the number of binucleated cells containing micronuclei, when compared to the numbers found in the concurrent control cultures (Table 4). However, the three higher concentrations analysed did not show an increase in the number of binucleated cells containing micronuclei. Hence, the response was not dose related. In addition, the response observed for this concentration was well within the historical range and the number of micronuclei found in the concurrent solvent control was slightly below the lower end of the historical data. Therefore, this result is considered to be a matter of chance and not biological relevant.
Table 1: Experiment 1 – Pulse treatment method with metabolic activation (S9-mix)
Treatm/recovery time (h) |
Dose level (µg/mL) |
Cell stage analysis/500 (MO-BN-MU) |
BN (%) |
CBPI |
CBPI (mean) |
RI (%) |
% Cytotox. (100-RI) |
Selected for MN analysis (+/-) |
MNBN/1000BN |
MNBN/2000BN (%) |
Statistics 1) (p-value) |
4/20 (+S9) |
NC |
227 267 6 |
53.40 |
1.56 |
1.55 |
100 |
0 |
+ |
6/1000 |
13/2000 |
- |
|
233 261 6 |
52.20 |
1.55 |
|
|
|
|
7/1000 |
0.65 |
|
|
1266 |
332 168 0 |
33.60 |
1.34 |
1.31 |
57 |
43 |
+ |
13/1000 |
21/2000 |
0.0841 |
|
|
357 142 1 |
28.40 |
1.29 |
|
|
|
|
8/1000 |
1.05 |
|
|
633 |
265 232 3 |
46.40 |
1.48 |
1.47 |
86 |
14 |
+ |
9/1000 |
13/2000 |
n.s. |
|
|
270 224 6 |
44.80 |
1.47 |
|
|
|
|
4/1000 |
0.65 |
|
|
317 |
260 234 6 |
46.80 |
1.49 |
1.51 |
91 |
9 |
+ |
9/1000 |
15/2000 |
n.s. |
|
|
245 251 4 |
50.20 |
1.52 |
|
|
|
|
6/1000 |
0.75 |
|
|
158 |
240 254 6 |
50.80 |
1.53 |
1.55 |
100 |
0 |
- |
- |
- |
- |
|
|
221 272 7 |
54.40 |
1.57 |
|
|
|
|
- |
|
|
|
79 |
255 243 5 |
48.31 |
1.50 |
1.52 |
94 |
6 |
- |
- |
- |
- |
|
|
242 250 8 |
50.00 |
1.53 |
|
|
|
|
- |
|
|
|
40 |
264 231 5 |
46.20 |
1.48 |
1.50 |
90 |
10 |
- |
- |
- |
- |
|
|
253 240 7 |
48.00 |
1.51 |
|
|
|
|
- |
|
|
|
CP25 |
372 127 1 |
25.40 |
1.26 |
1.27 |
48 |
52 |
+ |
32/1000 |
66/2000 |
<0.0001 |
|
|
363 136 1 |
27.20 |
1.28 |
|
|
|
|
34/1000 |
3.30 |
*** |
The fixed cells of the other dose levels (2.5 to 20 µg/mL) were stored without slide preparation
Table 2: Experiment 1 – Pulse treatment method without metabolic activation (S9-mix)
Treatm/recovery time (h) |
Dose level (µg/mL) |
Cell stage analysis/500 (MO-BN-MU) |
BN (%) |
CBPI |
CBPI (mean) |
RI (%) |
% Cytotox. (100-RI) |
Selected for MN analysis (+/-) |
MNBN/1000BN |
MNBN/2000BN (%) |
Statistics 1) (p-value) |
4/20 (-S9) |
NC |
224 267 9 |
53.40 |
1.57 |
1.56 |
100 |
0 |
+ |
6/1000 |
12/2000 |
- |
|
232 259 9 |
51.80 |
1.55 |
|
|
|
|
6/1000 |
(0.60) |
|
|
1266 |
348 149 3 |
29.80 |
1.31 |
1.32 |
56 |
44 |
+ |
11/1000 |
16/2000 |
n.s |
|
|
344 156 3 |
31.01 |
1.32 |
|
|
|
|
5/1000 |
(0.80) |
|
|
633 |
229 265 6 |
53.00 |
1.55 |
1.55 |
98 |
2 |
+ |
3/1000 |
10/2000 |
n.s. |
|
|
237 255 8 |
51.00 |
1.54 |
|
|
|
|
7/1000 |
(0.50) |
|
|
317 |
224 269 7 |
53.80 |
1.57 |
1.55 |
98 |
2 |
+ |
6/1000 |
12/2000 |
n.s. |
|
|
239 256 5 |
51.20 |
1.53 |
|
|
|
|
6/1000 |
(0.60) |
|
|
158 |
251 239 10 |
47.80 |
1.52 |
1.51 |
91 |
9 |
- |
- |
- |
- |
|
|
254 242 4 |
48.40 |
1.50 |
|
|
|
|
- |
|
|
|
79 |
245 251 4 |
50.20 |
1.52 |
1.49 |
88 |
12 |
- |
- |
- |
- |
|
|
277 218 10 |
43.17 |
1.47 |
|
|
|
|
- |
|
|
|
40 |
231 259 10 |
51.80 |
1.56 |
1.55 |
98 |
2 |
- |
- |
- |
- |
|
|
234 258 8 |
51.60 |
1.55 |
|
|
|
|
- |
|
|
The fixed cells of the other dose levels (2.5 to 20 µg/mL) were stored without slide preparation
Results to be continued in section "Overall remarks, attachements"
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
weight of evidence approach based on in vitro data and QSAR predictions
Link to relevant study records
- Endpoint:
- genetic toxicity in vivo, other
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- other:
- Justification for type of information:
- WoE approach determines a negative result overall for in vitro mutagenicity. It was concluded that the positive effects seen in the Chromosomal aberration test (1997) were generated at highly cytotoxic doses and should be interpreted with caution. A micronucleus study (2015) was conducted using an appropriate concentration range and with the determination of the cytotoxicity simultaneously during the main study. The results of this study were negative. These results were supported by three QSAR studies which all had negative results for genotoxicity.
- Reason / purpose for cross-reference:
- data waiving: supporting information
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Additional information from genetic toxicity in vitro:
Four in vitro genetic toxicity studies are available for the mutagenicity endpoint of MODA and were used for the assessment:
One in vitro gene mutation study in bacteria (Kazuhiko Saigo, 1997;equivalent/similar to OECD 471), one in vitro gene mutation study in mammalian cells (TÜV SÜD, 2012;OECD 476), one chromosome aberration test (Kazuhiko Saigo, 1997;equivalent/similar to OECD 473) with Chinese hamster lung (CHL)/IU) cells and one in vitro micronucleus study (B. Usta, BSc, 2015;OECD 487) with cultured human lymphocytes. Additionally, three QSAR calculations were conducted (OECD Toolbox, Vega and Test).
An in vitro gene mutation study in bacteria (Kazuhiko Saigo,1997) was conducted using five bacterial strains (Salmonella typhimurium TA98, TA100, TA1535 and TA1537 and E. coli WP2urvA) to evaluate the mutagenic potential of the substance. Six different doses, 156, 313, 625, 1250, 2500, and 5000µg/plate, were used in the main study with and without metabolic activation. Cytotoxicity was observed at concentrations of 2500 and 5000 µg/plate. In this test, the substance did not show mutagenic potential with and without metabolic activation.
In an in vitro mammalian cell gene mutation study using mouse lymphoma L5178Y cells (TÜV SÜD, 2012; OECD 476) cytotoxic effects (cell growth inhibition) were observed at 2.5µg/mL and above concentrations in a pretest. Concentrations of 2.5µg/mL, 1.25µg/mL, 0.625µg/mL and 0.3125µg/mL were used for the main test. In this test the substance was determined to be non-mutagenic to the L5178Y TK+/- clone, both in the absence and presence of metabolic activation.
In the chromosome aberration study using cultured Chinese hamster lung-derived fibroblast cells (Kazuhiko Saigo, 1997) a positive result was concluded for the substance. The test was carried out with 12.5, 25, 50, and 100 µg/mL in the absence of metabolic activation with the treatment periods of 24 and 48 hrs, and with 44.4, 66.7, 100, and 150 µg/mL in the presence of metabolic activation without S9 Mix, and at 444, 667, 1000, and 1500 µg/mL in the presence of metabolic activation with S9 Mix. Doses were set so that the cell survival rate ranged between 20% and 80% based on the results of a range finding study. Cytotoxicity was not evaluated during the main study. Positive results were observed at concentrations of 50 and 100 µg/mL in the absence of metabolic activation with the treatment periods of 24 and 48 hrs. Both concentrations were above the 50% cell growth inhibition concentration (GI50) of around 48 and 35 µg/mL, respectively, as determined in the pretest. With metabolic activation a positive result was observed with S9 Mix at 1000 µg/mL. Here, a GI50 of 610µg/mL was calculated from the range finding study. With metabolic activation without S9 Mix positive results were observed at all four concentrations (44.4, 66.7, 100, and 150 µg/mL). For this method, a GI50 of 80µg/mL was calculated from the range finding study. As most of the positive responses were observed at doses above the GI50 value the study is considered of limited value because the observed chromosome damage may be induced as a secondary effect of cytotoxicity (ECHA guidance R7a, 2015 section R.7.7.4.1). Also cytotoxicity was not determined in the main study but in a separate test.
As the positive result of the chromosome aberration test (equivalent/similar to OECD 473) with Chinese hamster lung cells was considered to be questionable a follow up study was carried out to determine the potential for chromosome damaging of the substance. Due to the observed high cytotoxicity in the first chromosome aberration test using Chinese hamster lung cells, the follow up test was conducted with cultured human lymphocytes in a micronucleus test according to OECD 487 (B. Usta, BSc, 2015) with the following concentrations: Experiment 1: 1266, 633, 317, 158, 79, 40, 20, 10, 5, 2.5 µg/mL (pulse treatment method with and without metabolic activation; Experiment 2: 1266, 886, 620, 434, 303, 213 µg/mL (pulse treatment method without metabolic activation); Experiment 2: 1266, 886, 620, 434, 303, 213, 149, 104, 72.8, 51.1, 35.8, 25.0, 17.5, 12.3 µg/mL (continuous treatment method without metabolic activation). In this test the Cytotoxicity was determined from the Cytokinesis-Block Proliferation Index (CBPI) and three test concentrations (not including the solvent and positive controls) that meet the acceptability criteria (appropriate cytotoxicity, number of cells, etc) could be evaluated as required according to the test guideline. From the results obtained in this in vitro micronucleus test it was concluded that, under the reliable conditions used in this study, the test substance MODA was not clastogenic and/or aneugenic to cultured human lymphocytes.
In addition three independent QSAR studies were conducted using different models. All three models predict that primary amines do not have a genotoxic potential.
The available data were used in a weight of evidence approach to determine the potential of the substance for genetic toxicity. It was concluded that the substance has no mutagenic potential and further in vivo testing was not deemed necessary.
Justification for selection of genetic toxicity endpoint
The mammalian cell micronucleus test (OECD 487) is the most recent study.
Justification for classification or non-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.
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