Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Link to relevant study records

Referenceopen allclose all

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
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Link to relevant study records

Referenceopen allclose all

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.

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)