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EC number: 204-539-4 | CAS number: 122-39-4
- 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
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:
- supporting study
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- other: unsuitable test system
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- Ames test modified
- 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
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- S. typhimurium TA 1538
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- S. typhimurium, other: G46, C3076, D3052
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- E. coli WP2
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- E. coli, other: WP uvrA-
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- liver fraction (S9) from Aroclor 1254 pretreated rats
- Test concentrations with justification for top dose:
- 10000-fold concentration range for the test, 10-fold concentration range per plate
- Species / strain:
- other: all
- 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
- Conclusions:
- Interpretation of results (migrated information):
negative
Under the conditions of the study, diphenylamine was found non mutagenic/carcinogenic in the Modified Ames Test - 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:
- other: Comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- not specified
- 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
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- S. typhimurium TA 97
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S-9 (9,000 g supernatant) fractions of Aroclor 1254-induced male Sprague-Dawley rat and male Syrian hamsters livers
- Test concentrations with justification for top dose:
- 0, 1, 3, 10, 33, 66, 100, 166, 333 µg/plate
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: [distilled water, DMSO, 95% ethanol and acetone]
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- concurrent solvent
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- without metabolic activation, strains TA1535 and TA100
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- concurrent solvent
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- withoutf metabolic activation, strains TA97 and TA1537
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- concurrent solvent
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 4-nitro-o-phenylenediamine
- Remarks:
- without metabolic activation, strain TA98
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- concurrent solvent
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- with metabolic activation, all strains
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: preincubation
DURATION
- Preincubation period: 20 min
- Exposure duration: 2 days
DETERMINATION OF CYTOTOXICITY
- Method: other: density of the background lawn and evolution of hte number of his+ colonies - Evaluation criteria:
- A chemical is judged mutagenic (+) or weakly mutagenic (+W) if it produces a reproducible dose-related response over the solvent control in replicate trials
A chemical is questionable (?) if results of individual trials are not reproducible, if increases in his+ revertants do not meet the criteria for "+W" or if only single doses produce icnreases in his+ revertants
A chemical is non mutagenic if it does not meet the criteria for questionable or mutagenic response - Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- other: only values available
- Species / strain:
- S. typhimurium TA 97
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- other: only values available
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- other: only values available
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- other: only values available
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not specified
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- other: only values available
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- negative
Under the conditions of the study, diphenylamine was found non mutagenic - Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable tu guideline study with acceptable restrictions: deviations were not considered to have affected the scientific validity and interpretation of the results
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- cells exposed to the test substance for 22 hours at some concentrations in the absence of metabolic activation
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Preliminary toxicity test: with activation: 12.5 to 150 µg/ml; without activation: 20 to 5000 µg/ml
Mutagenicity test: with activation: 12.5, 25, 50, 55, 60, 65 µg/ml; without activation: 10, 20, 40, 50, 60, 70, 80 µg/ml - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: [DMSO]
- Untreated negative controls:
- yes
- Remarks:
- vehicle
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO (concentration: 1%)
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- with metabolic activation
- Untreated negative controls:
- yes
- Remarks:
- vehicle
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO (concentration: 1%)
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- without metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in suspension
DURATION
- Exposure duration: 22 hours
- Expression time (cells in growth medium): 2 hours (accumation of metaphase cells)
- Fixation time (start of exposure up to fixation or harvest of cells): 24 hours
SPINDLE INHIBITOR (cytogenetic assays): colcemid
NUMBER OF CELLS EVALUATED: 100 cells scored per dose
DETERMINATION OF CYTOTOXICITY
- Method: other: presence of metaphase cells, reduced cell counts
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreplication: yes
- Other: aneuploidy - Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- toxicity and no metaphase cells observed: 80 µg/ml; reduced cell counts: 70 µg/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- toxicity an no metaphase cells observed: 50-150 µg/ml; reduced cell counts: 55 µg/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
- Conclusions:
- Interpretation of results (migrated information):
negative without metabolic activation
positive with metabolic activation
DPA Super-Refined diphenylamine was a weak clastogen when tested with Chinese hamster ovary cells in vitro in the presence of S9 mix, at concentrations deemed toxic to the cells - Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions: nonactivation study (S9-) was not conducted
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Type of assay:
- mammalian cell gene mutation assay
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with
- Metabolic activation system:
- Aroclor 1254-induced and non induced S9 fractions from male Sprague-Dawley rat liver
- Test concentrations with justification for top dose:
- 0, 3.79, 5.06, 6.75, 9.00 (x 10^-5) M
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: [normal saline, media, DMSO or ethanol (for diphenylamine)]
- Justification for choice of solvent/vehicle: - - Untreated negative controls:
- yes
- Remarks:
- untreated media
- Negative solvent / vehicle controls:
- yes
- Remarks:
- ethanol final concentration: 1%
- True negative controls:
- not specified
- Positive controls:
- not specified
- Positive control substance:
- not specified
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in suspension
DURATION
- Preincubation period:-
- Exposure duration: 3 hours
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): 7 days
- Fixation time (start of exposure up to fixation or harvest of cells):no data
SELECTION AGENT (mutation assays): trifluorothymidine (TFT)
SPINDLE INHIBITOR (cytogenetic assays):-
STAIN (for cytogenetic assays):-
NUMBER OF REPLICATIONS: 3 replicates per concentration per chemical
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency, cell survival - Statistics:
- Parameters: number of mutants per plate, X: number of titrations used for each concentration, Y: number of concentrations with better than 20% survival for each compound
Methods: linear regression, t-test and a Monte Carlo comparison of the 2 statistical methods - Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- not specified
- Conclusions:
- Interpretation of results (migrated information):
negative with metabolic activation
Under the conditions of the study diphenylamine was found non mutagenic - 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:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- other: EPA 84-1
- Deviations:
- yes
- Remarks:
- actual concentration of methyl methanesulphonate not given
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- yes
- Remarks:
- actual concentration of methyl methanesulphonate not given
- Principles of method if other than guideline:
- actual concentration of methyl methanesulphonate not given
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- self certified
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- TK
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Additional strain / cell type characteristics:
- other: substrain 3.7.2c
- Metabolic activation:
- with and without
- Metabolic activation system:
- liver microsomes preparations (S9) from Aroclor 1254 induced SD rats
- Test concentrations with justification for top dose:
- Test 1 without activation: 5, 10, 20, 40, 50, 60, 80 µg/ml
Test 1 with activation: 5, 10, 20, 40, 50, 60, 80 µg/ml
Test 2 with activation: 5, 10, 20, 40, 50, 60, 70 µg/ml - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: [DMSO (final concentration 1%)]
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- methylmethanesulfonate
- Remarks:
- without activation Migrated to IUCLID6: 10, 15 nl/ml
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 3-methylcholanthrene
- Remarks:
- with activation Migrated to IUCLID6: 2.0-4.0 µg/ml
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in suspension
DURATION
- Preincubation period: no data
- Exposure duration: 4 hours
- Expression time (cells in growth medium): no data
- Selection time (if incubation with a selection agent): 10 to 14 days
- Fixation time (start of exposure up to fixation or harvest of cells): no data
SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays):
STAIN (for cytogenetic assays):
NUMBER OF REPLICATIONS:-
NUMBER OF CELLS EVALUATED: 3 x10^6 (total sample for the determination of cloning efficiency)
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth - Evaluation criteria:
- Without activation: a mutagenic response must exceed minimum criterion of 130.5 x 10^-6 mutant frequency
With activation: a mutagenic response must exceed minimum criterion of 138.2 x 10^-6 (Test 1) and 101.0 x 10^-6 (Test 2) mutant frequency - Key result
- Species / strain:
- not specified
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- reduction in cell growth at 7.81 µg/ml; total cell killing at and above 125 µg/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Key result
- Species / strain:
- not specified
- Metabolic activation:
- with
- Genotoxicity:
- positive
- Remarks:
- 40 and 80 µg/ml (Test 1); 20, 30, 40, 50 and 70 µg/ml (Test 2)
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- reduction in cell growth at 7.81 µg/ml; total cell killing at and above 125 µg/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results (migrated information):
negative without metabolic activation
positive with metabolic activation
Under the conditions of testing diphenylamine was inactive in the mouse lymphoma forward mutation assay without S9 metabolic activation and weakly active in the presence of S9 metabolic activation - Endpoint:
- in vitro DNA damage and/or repair study
- Remarks:
- Type of genotoxicity: DNA damage and/or repair
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- other: unsuitable test system
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 482 (Genetic Toxicology: DNA Damage and Repair, Unscheduled DNA Synthesis in Mammalian Cells In Vitro)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Type of assay:
- DNA damage and repair assay, unscheduled DNA synthesis in mammalian cells in vitro
- Species / strain / cell type:
- hepatocytes:
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- not applicable
- Test concentrations with justification for top dose:
- 0.5, 1, 5, 10, 50, 100, 500, 1000 nmoles/ml
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: [DMSO]
- Justification for choice of solvent/vehicle:- - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 2-acetylaminofluorene
- Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- N-ethyl-N-nitro-N-nitrosoguanidine
- Evaluation criteria:
- A positive response to UDS is induced when at least 2 successive concentrations produced nuclear grain counts which exceeded those of the control by at least 2 standard deviations of the control value
- Species / strain:
- hepatocytes: Fisher adult rat
- Metabolic activation:
- not applicable
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- >=100 nmoles/ml
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results (migrated information):
negative
Under the conditions of the study, diphenylamine was found non mutagenic/carcinogenic in Hepatocytes UDS - Endpoint:
- in vitro DNA damage and/or repair study
- Remarks:
- Type of genotoxicity: DNA damage and/or repair
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 479 (Genetic Toxicology: In Vitro Sister Chromatid Exchange Assay in Mammalian Cells)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Type of assay:
- sister chromatid exchange assay in mammalian cells
- Species / strain / cell type:
- lymphocytes: human adult non-smoking subjects
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 or phenobarbital and benzoflavone induced microsmola preparation
- Test concentrations with justification for top dose:
- 0.6, 6, 60 µg/ml culture (respectively 3.5 x 10^-6, 3.5 x10^-5, 3 x 10^-4 M)
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: [DMSO]
- Justification for choice of solvent/vehicle:- - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- without metabolic activation
Migrated to IUCLID6: 0.01 µg/ml culture - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- not specified
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- with metabolic activation
Migrated to IUCLID6: 10^-5 M - Details on test system and experimental conditions:
- DURATION
- Exposure duration: 48 hours (Protocole 1) and 4 hours (Protocoles 2,3)
- Fixation time (start of exposure up to fixation or harvest of cells): 72 hours
SPINDLE INHIBITOR (cytogenetic assays): Colcemid (Gibco)
STAIN (for cytogenetic assays): 5-bromodeoxyuridine
DETERMINATION OF CYTOTOXICITY
- Method: other: proliferating rate index - Statistics:
- Methods: randomized block Anova 2 on transformed data; Dunnet test for the comparison among treatments; chi-square test
Parameters: SCE frequencies; cell kinetics - Metabolic activation:
- without
- Genotoxicity:
- positive
- Remarks:
- only in protocol 1 at the higher concentration
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- not applicable
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results (migrated information):
positive without metabolic activation
The study suggests that diphenylamine is a weak direct genotoxic agent in this assay - 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:
- other: Guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- EPA OPP 84-2
- Deviations:
- yes
- Remarks:
- bacterial strains capable of detecting oxidising mutagens, cross-linking agents and hydrazines not tested
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- no data concerning the results of negative and positive controls, neither cytotoxicity
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Additional strain / cell type characteristics:
- not specified
- Species / strain / cell type:
- S. typhimurium TA 1538
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- With S9: 10.0, 33.3, 66.7, 100, 333, 667 µg/plate
Without S9: 6.67, 10.0, 33.3, 66.7, 100, 333 µg/plate - Untreated negative controls:
- yes
- Remarks:
- solvent
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene 2.5 µg/plate
- Remarks:
- with activation, for all strains
- Untreated negative controls:
- yes
- Remarks:
- solvent
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- Remarks:
- without activation, for strains TA98, TA1538
Migrated to IUCLID6: 1.0 µg/plate - Untreated negative controls:
- yes
- Remarks:
- solvent
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- without activation, for strains TA100, TA1535
Migrated to IUCLID6: 2.0 µg/plate - Untreated negative controls:
- not specified
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- not specified
- Positive controls:
- yes
- Positive control substance:
- other: ICR-191 2.0 µg/plate
- Remarks:
- without activation, for strain TA1537
- Details on test system and experimental conditions:
- DURATION
- Exposure duration: 48 +/- 8 hours
NUMBER OF CELLS EVALUATED: 0.5 x 10^9 - Evaluation criteria:
- TA98, TA100: A test substance was considered positive if the mean number of revertants were twice the control result
TA1535, TA1537, TA1538: A 3-fold increases is required to register a positive result - Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Species / strain:
- S. typhimurium TA 1538
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- not determined
- Vehicle controls validity:
- not specified
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- not specified
- Conclusions:
- Interpretation of results (migrated information):
negative
Under the conditions of the study diphenylamine did not induce reverse gene mutation in the selected bacterial tester strain
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Link to relevant study records
- Endpoint:
- in vivo mammalian cell study: DNA damage and/or repair
- Remarks:
- Type of genotoxicity: DNA damage and/or repair
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 486 (Unscheduled DNA Synthesis (UDS) Test with Mammalian Liver Cells in vivo)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- self certified
- Type of assay:
- unscheduled DNA synthesis
- Species:
- rat
- Strain:
- Fischer 344
- Sex:
- male
- Route of administration:
- oral: unspecified
- Duration of treatment / exposure:
- 2-4 hour and 15-16 hour after dosing
- Frequency of treatment:
- Single dose
- Post exposure period:
- None
- Remarks:
- Doses / Concentrations:
200, 500, 800, 1500 & 2000 mg/kg bw
Basis:
no data
(range finding test) - Remarks:
- Doses / Concentrations:
0, 750, 1500 & 2000 mg/kg bw
Basis:
no data
(UDS test) - No. of animals per sex per dose:
- 4 rats/sex/dose/timepoint
- Control animals:
- yes, concurrent vehicle
- Positive control(s):
- N-dimethylnitrosamine
- Route of administration: IP injection
- Doses / concentrations: 10 & 15 mg/kg bw, for the 2-to 4- hour and 15- to 16- hour timepoints, respectively - Tissues and cell types examined:
- Liver: hepatocytes
- Details of tissue and slide preparation:
- DETAILS OF SLIDE PREPARATION:
Hepatocytes were obtained by mechanicle dispersion of excised liver tissue, centrifuged and re-suspended to perform a viable cell count
Isolated cells were re-suspended and replicate cultures prepared, then incubated and radiolabelled with high specific activity thymidine for 4 hrs
Cells were washed, fixed and allowed to dry, then mounted on glass slides and dipped in an autoradiogaphic emulsion
METHOD OF ANALYSIS:
Autoradiography - Sex:
- male
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- slight effects at the highest doses tested
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- Interpretation of results (migrated information): negative
Diphenylamine was found inactive under the conditions of the study - Executive summary:
Fisher 344 rats were orally administered diphenylamine at : 0, 750, 1500 & 2000 mg/kg bw for the 2 -4 hr and the 15 -16 hr timepoints. All animals were observed throughout the duration of the assay for toxic symptoms and mortality. Moreover, hepatocytes were examined by autoradiography and the mean net nuclear grain count was determined from triplicates coverslips for each treated animal. The results were negative.
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Deviations:
- yes
- Remarks:
- the frequency of PCEs vs NCEs was determined by scoring the frequency of both observed in the optic fiels while scoring the first 1000 erythrocytes
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- self certified
- Type of assay:
- endogenous gene animal assay
- Species:
- mouse
- Strain:
- ICR
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Age at study initiation: 8 weeks - Route of administration:
- oral: unspecified
- Vehicle:
- - Vehicle(s)/solvent(s) used: [corn oil]
- Duration of treatment / exposure:
- 72 h
- Frequency of treatment:
- Single dose
- Post exposure period:
- Test substance: 72 h
Negative/Positive controls: 24 h - Remarks:
- Doses / Concentrations:
250, 500 & 1000 mg/kg
Basis:
no data
males - Remarks:
- Doses / Concentrations:
375, 750 & 1500 mg/kg
Basis:
no data
females - No. of animals per sex per dose:
- 5 mice/sex/dose
- Control animals:
- yes
- Positive control(s):
- cyclophosphamide
- Route of administration: oral
- Doses / concentrations: 80 mg/kg - Tissues and cell types examined:
- Bone marrow erythrocytes
- Evaluation criteria:
- A positive response is indicated by a statistically increase in the incidence of micronucleated polychromatic erythrocytes compared with the vehicle control
Bone marrow toxicity is indicated by a statistically significant decrease in the ratio of polychromatic to normochromatic erythrocytes - Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- no effects
- Vehicle controls validity:
- valid
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): no significant increases
- Ratio of PCE/NCE (for Micronucleus assay): range of 0.2-0.49 (males) and 0.34-0.79 (females)
- Appropriateness of dose levels and route: dose levels determined in a previous range finding study - Conclusions:
- Interpretation of results (migrated information): negative
Under the conditions of the in vivo study, diphenylamine did not induce chromosomal damage in mouse erythrocytes - Executive summary:
Negative results up to lethal oral gavage doses from an mouse micronucleus assay were reported. ICR mice were treated by single oral gavage of 250, 500 or 1000 mg/kg bw (males) or 375, 750 or 1500 mg/kg bw (females) diphenylamine (99.9%) and frequencies of micronucleated polychromatic erythrocytes in bone marrow cells were investigated 24, 48 or 72 hours after treatment.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
DPA was negative in two Salmonella gene mutation tests. In conclusion the whole amount of data indicates that diphenylamine may not be mutagenic in humans.
Short description of key information:
In vitro gene mutation study in bacteria: Non mutagenic (Zeiger, 1988; Probst, 1981)
Justification for classification or non-classification
Based on the availablee study data for the substance, diphenylamine is notanticipated to be mutagenic in accordance with the criteria specified in Regulation 1272/2008 (CLP)
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