4,4'-methylenebis[N,N-bis(2,3-epoxypropyl)aniline]

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / micronucleus study

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No guideline cited but according to published method

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1984

Report date:

1984

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Method according to Clive, D. and Spector, J. (1975). Laboratory procedure for assessing specific locus mutations at the TK locus in cultures L5178Ylymphoma cells. Mutation Research 31, 17 - 29.

GLP compliance:

yes

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-Methylenedianiline, oligomeric reaction products with 1-chloro-2,3-epoxypropane

EC Number:

500-062-3

EC Name:

4,4'-Methylenedianiline, oligomeric reaction products with 1-chloro-2,3-epoxypropane

Cas Number:

28390-91-2

IUPAC Name:

4-[(4-aminophenyl)methyl]aniline; 2-(chloromethyl)oxirane

Test material form:

liquid: viscous

Details on test material:

Batch no. 4356

Method

Target gene:

TK -locus

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

heterozygous subline , TK +/- obtained from Dr. D. Clive

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 from Aroclor induced rat livers

Test concentrations with justification for top dose:

without metabolic activation: 0, 1.28, 2.56, 5.12, 10.25, 20.5 micrograms/ml
with metabolic activation: 0, 4.17, 8.33, 16.67, 33.35, 66.7 microgram/ml

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

Migrated to IUCLID6: 0.75 microgram/ml

Details on test system and experimental conditions:

Preliminary cytotoxicity test: highest concentration for mutagenicity assay causes 85% growth reduction after 4-hour treatment and 72 -hour suspension growth. Cells were grown in Fishers medium F10, with 10% horse serum. Final DMSO concentration was always < 1%.
Cells were exposed for 4 hours at seven concentrations of 15.63 - 1000 micrograms/ml. Total growth inhibition (cytotoxicity) achieved at 250 micrograms/ml within 48 hours.
Main experiment: 3 x 105 cells were seeded per culture and were treated for 4 hours with TGMDA in the presence or absence of S9 activating liver enzymes, followed by a growth period of 3 days without exposure. After this expression phase cells were plated in soft-agar in petri-dishes containing 50 micrograms/ml of BUdR, and additional 14 days of incubation followed. With a colony counter the colonies were counted .
The results are expressed as number of induced TK-/- mutants/106 cells.

Evaluation criteria:

To be a positive result, the colony count of the treated plates has to be 2.5 times higher than the solvent control.

Statistics:

no statistical methods were applied

Results and discussion

Test results

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

3 tests without metabolic activation and one test with metabolic activation was performed. In all experiments the highest concentration yielded the highest amount of mutant colonies, but at growth rates of 0.3 - 1.7 % (or cytotoxicity of >98%). (see tables)
Nevertheless, the result was consitant throughout the three experiments and was judged to be positive.

Remarks on result:

other: strain/cell type: not determined

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive

Under the conditions of the test in mouse lymphoma cells, the product TK 10884 was mutagenic with and without S9-activating enzymes.

Executive summary:

Under the conditions of the test in mouse lymphoma cells, the product TK 10884 was mutagenic with and without S9-activating enzymes.